Introduction:Crude oil-contaminated soils harbor bacteria with specialized hydrocarbon-degrading capabilities, yet the Stenotrophomonas genus remains incompletely characterized at the species level. Strain ASS1, a motile Gram-negative rod, was isolated from crude oil-contaminated soil in Tabasco, Mexico, and initial 16S rRNA gene sequencing indicated affiliation with Stenotrophomonas, warranting full polyphasic characterization. Methods:The complete genome was sequenced using Illumina MiSeq and assembled into a single closed contig (4,564,481 bp; G+C content 66.59%; GenBank: CP031167). Genome quality was assessed with BUSCO v6.0.0 and CheckM2. Species boundaries were evaluated using average nucleotide identity (ANIb via JSpeciesWS) and digital DNA-DNA hybridization (dDDH via GGDC Formula 2) against 38 representative Stenotrophomonas genomes. Taxonomic placement was further resolved by TYGS, MLST, and core genome phylogenomics. PAH catabolic capacity was assessed through genome annotation and in vitro growth assays on four PAHs as sole carbon sources, analyzed by Kruskal-Wallis test. Results:BUSCO completeness was 99.7% and CheckM2 confirmed 100.0% completeness with 0.0% contamination. Despite 99.86% 16S rRNA gene identity to Stenotrophomonas geniculata ATCC 19374T, ANIb against the closest neighbor Stenotrophomonas riyadhensis CFS3442T was 93.30%, and dDDH was 50.4% (95% CI: 47.8-53.1%), both below established species thresholds. No pairwise comparison across the 38 genomes exceeded 54.3% dDDH. TYGS, MLST, and core genome phylogenomics consistently resolved ASS1 as a distinct lineage. Pan genome analysis confirmed an open pan genome (alpha = 0.485). A complete upper pathway PAH catabolic gene cassette was identified, and in vitro growth was statistically equivalent across all four PAH substrates (Kruskal-Wallis H(3) = 2.403, p = 0.493). Discussion:Collectively, the genomic, phylogenomic, and phenotypic evidence supports ASS1 as the type strain of a novel species, Stenotrophomonas oleivorans sp. nov. (= WFCC 1006/CM-CNRG 934T), with demonstrated broad spectrum PAH degradation capacity relevant to bioremediation.

Acinetobacter baumannii has emerged as a critical global health threat due to its exceptional survival skills in adverse environment and its ability to acquire antibiotic resistance, presenting significant challenges for infection treatment and control. The World Health Organization has classified carbapenem-resistant A. baumannii as a “Critical Priority” pathogen to guide research and the development of control and prevention strategies. Epidemiological surveillance methodologies provide the tools necessary for classifying A. baumannii into international clonal lineages, facilitating the analysis of molecular characteristics, global dissemination, and evolution. This study provides a detailed analysis of the molecular epidemiology of A. baumannii in Mexico, focusing on identifying the main international clonal lineages. Genomic analyses of 146 genomes, along with information from previous studies, identified 24 different sequence types according to the Oxford Scheme. The major international clone IC2 (CC208) was identified and harbors β-lactamases OXA-66, ADC-30, OXA-72, and is predicted to possess the OCL1 locus. The international clone IC5 (CC205) carries β-lactamase OXA-65, along with ADC-214 and OXA-239, with OCL10 predicted in 82.2% of the genomes. The international clone IC7 (CC229) harbors β-lactamase OXA-64, as well as ADC-174 and ADC-214, with OCL6 and OCL7 loci predicted. These international clones were identified in different periods and regions of Mexico and are likely to be widely distributed throughout the country. The analysis of each lineage reveals distinct molecular characteristics, including sequence types, capsule typing, outer core loci, and specific antibiotic resistance profiles. Understanding these features is crucial for elucidating their roles in infection dynamics, resistance mechanisms, and their impact on clinical outcomes.

Rhizobial phosphatidylcholine (PC) is thought to be a critical phospholipid for the symbiotic relationship between rhizobia and legume host plants. A PC-deficient mutant of Sinorhizobium meliloti overproduces succinoglycan, is unable to swim, and lacks the ability to form nodules on alfalfa (Medicago sativa) host roots. Suppressor mutants had been obtained which did not overproduce succinoglycan and regained the ability to swim. Previously, we showed that point mutations leading to altered ExoS proteins can reverse the succinoglycan and swimming phenotypes of a PC-deficient mutant. Here, we report that other point mutations leading to altered ExoS, ChvI, FabA, or RpoH1 proteins also revert the succinoglycan and swimming phenotypes of PC-deficient mutants. Notably, the suppressor mutants also restore the ability to form nodule organs on alfalfa roots. However, nodules generated by these suppressor mutants express only low levels of an early nodulin, do not induce leghemoglobin transcript accumulation, thus remain white, and are unable to fix nitrogen. Among these suppressor mutants, we detected a reduced function mutant of the 3-hydoxydecanoyl-acyl carrier protein dehydratase FabA that produces reduced amounts of unsaturated and increased amounts of shorter chain fatty acids. This alteration of fatty acid composition probably affects lipid packing thereby partially compensating for the previous loss of PC and contributing to the restoration of membrane homeostasis.

Computational modeling (CM) is a versatile scientific methodology used to examine the properties and behavior of complex systems, such as polymeric materials for biomedical bioengineering. CM has emerged as a primary tool for predicting, setting up, and interpreting experimental results. Integrating in silico and in vitro experiments accelerates scientific advancements, yielding quicker results at a reduced cost. While CM is a mature discipline, its use in biomedical engineering for biopolymer materials has only recently gained prominence. In biopolymer biomedical engineering, CM focuses on three key research areas: (A) Computer-aided design (CAD/CAM) utilizes specialized software to design and model biopolymers for various biomedical applications. This technology allows researchers to create precise three-dimensional models of biopolymers, taking into account their chemical, structural, and functional properties. These models can be used to enhance the structure of biopolymers and improve their effectiveness in specific medical applications. (B) Finite element analysis, a computational technique used to analyze and solve problems in engineering and physics. This approach divides the physical domain into small finite elements with simple geometric shapes. This computational technique enables the study and understanding of the mechanical and structural behavior of biopolymers in biomedical environments. (C) Molecular dynamics (MD) simulations involve using advanced computational techniques to study the behavior of biopolymers at the molecular and atomic levels. These simulations are fundamental for better understanding biological processes at the molecular level. Studying the wide-ranging uses of MD simulations in biopolymers involves examining the structural, functional, and evolutionary aspects of biomolecular systems over time. MD simulations solve Newton’s equations of motion for all-atom systems, producing spatial trajectories for each atom. This provides valuable insights into properties such as water absorption on biopolymer surfaces and interactions with solid surfaces, which are crucial for assessing biomaterials. This review provides a comprehensive overview of the various applications of MD simulations in biopolymers. Additionally, it highlights the flexibility, robustness, and synergistic relationship between in silico and experimental techniques.

Infectious endophthalmitis is a severe ophthalmic emergency. This infection can be caused by bacteria and fungi. For efficient treatment, the administration of antimicrobial drugs to which the microbes are susceptible is essential. The aim of this study was to identify micro-organisms in biopsies of Mexican endophthalmitis patients using metagenomic next-generation sequencing and determine which antibiotic resistance genes were present in the biopsy samples. In this prospective case study, 19 endophthalmitis patients were recruited. Samples of vitreous or aqueous humour were extracted for DNA extraction for metagenomic next-generation sequencing. Analysis of the sequencing results revealed the presence of a wide variety of bacteria in the biopsies. Resistome analysis showed that homologues of antibiotic resistance genes were present in several biopsy samples. Genes possibly conferring resistance to ceftazidime and vancomycin were detected in addition to various genes encoding efflux pumps. Our findings contrast with the widespread opinion that only one or a few bacterial strains are present in the infected tissues of endophthalmitis patients. These diverse communities might host many of the resistance genes that were detected, which can further complicate the infections.

We report the draft genomes of four Kluyveromyces marxianus isolates obtained from the elaboration process of henequen (Agave fourcroydes) mezcal, a Mexican alcoholic beverage. The average nucleotide identity analysis revealed that isolates derived from agave plants are distinct from those from other environments, including agave fermentations.

We announce the genome sequencing, assembly, and annotation of the OLsAu1 strain and its taxonomic assignment to Pseudomonas yamanorum. The isolate comes from a wild edible ectomycorrhizal Lactarius sp. mushroom in the Abies forest. There is information regarding the strain's ability to promote plant growth, indicating its potential application in forestry.

Cyclospora cayetanensis is an enteric coccidian parasite responsible for gastrointestinal disease transmitted through contaminated food and water. It has been documented in several countries, mostly with low-socioeconomic levels, although major outbreaks have hit developed countries. Detection methods based on oocyst morphology, staining, and molecular testing have been developed. However, the current MLST panel offers an opportunity for enhancement, as amplification of all molecular markers remains unfeasible in the majority of samples. This study aims to address this challenge by evaluating two approaches for analyzing the genetic diversity of C. cayetanensis and identifying reliable markers for subtyping: core homologous genes and mitochondrial genome analysis. A pangenome was constructed using 36 complete genomes of C. cayetanensis , and a haplotype network and phylogenetic analysis were conducted using 33 mitochondrial genomes. Through the analysis of the pangenome, 47 potential markers were identified, emphasizing the need for more sequence data to achieve comprehensive characterization. Additionally, the analysis of mitochondrial genomes revealed 19 single-nucleotide variations that can serve as characteristic markers for subtyping this parasite. These findings not only contribute to the selection of molecular markers for C. cayetanensis subtyping, but they also drive the knowledge toward the potential development of a comprehensive genotyping method for this parasite.

The spread of ESBL-producing Escherichia coli has constantly increased in both clinical and community infections. Actually, the main ESBL reported is the CTX-M family, which is widely disseminated between the Enterobacteriaceae family. The epidemiology of the CTX-M family shows the CTX-M-15 variant dominating worldwide, followed by CTX-M-14 and CTX-M-27. The specific ESBL-producing E. coli clones included mainly the sequence types ST131, ST405, and ST648. In this report, we present the molecular characterization of ESBL-producing E. coli clinical isolates from eight hospitals in Mexico. From a collection of 66 isolates, 39 (59

Pediococcus pentosaceus 1101 was identified by using 16S rRNA and MALDI-Biotyper. The strain was exposed to conditions that resemble the gastrointestinal tract (GT) to evaluate its probiotic properties. That included the growth kinetics, proteolytic and inhibitory activities within a pH range, survival at low pH and in the presence of bile salts, antagonistic activity, cell-adhesion properties, and antibiotic resistance. The evaluation was followed by a genomic and proteomic analysis that involved the identification of proteins obtained under control and gastrointestinal conditions. The strain showed antagonistic activity against Gram-negative and Gram-positive bacteria, high resistance to acidity (87% logarithmic survival rate, pH 2) and bile salts (99% logarithmic survival rate, 0.5% w/v), and hydrophobic binding, as well as sensitivity to penicillin, amoxicillin, and chloramphenicol. On the other hand, P. pentosaceus 1101 has a genome size of 1.76 Mbp, with 1754 coding sequences, 55 rRNAs, and 33 tRNAs. The proteomic analysis showed that 120 proteins were involved in mechanisms in which the strain senses the effects of acid and bile salts. Moreover, the strain produces at least one lytic enzyme (N-acetylmuramoyl-L-alanine amidase; 32 kDa) that may be related to the antimicrobial activity. Therefore, proteins identified might be a key factor when it comes to the adaptation of P. pentosaceus 1101 into the GT and associated with its technological and probiotic properties.

Antimicrobial resistance is a major global public health problem, with fluoroquinolone-resistant strains of Escherichia coli posing a significant threat. This study examines the genetic characterization of ESBL-producing E. coli isolates in Mexican hospitals, which are resistant to both cephalosporins and fluoroquinolones. A total of 23 ESBL-producing E. coli isolates were found to be positive for the qepA gene, which confers resistance to fluoroquinolones. These isolates exhibited drug resistance phenotypes and belonged to specific sequence types and phylogenetic groups. The genetic context of the qepA gene was identified in a novel genetic context flanked by IS26 sequences. Mating experiments showed the co-transfer of qepA1 and chrA determinants alongside blaCTX-M-15 genes, emphasizing the potential for these genetic structures to spread among Enterobacterales. The emergence of multidrug-resistant Gram-negative bacteria carrying these resistance genes is a significant clinical concern for public healthcare systems.

We present the genome of a highly copper-tolerant pink-pigmented facultative methylotroph isolated from the rhizosphere of grasses growing close to mine tailings. Based on whole-genome taxonomic analyses, this isolate was named Methylobacterium radiotolerans MLP1. Studies are in progress to infer its genome-based copper resistome.

Babesia bovis, a tick-borne intraerythrocytic protozoan parasite that belongs to the phylum Apicomplexa, is one of the etiological agents of bovine babesiosis, a highly prevalent disease in tropical and subtropical countries that causes significant morbidity and deaths in cattle. This report presents the draft genome sequences of attenuated and virulent B. bovis strains of Mexican origin.

Verticillium dahliae is one of the most devastating soilborne pathogens for horticulture production. The pathogen has a broad host range and currently there is no effective chemical disease management, therefore, novel sustainable integrated disease management strategies should be considered. In this respect, we questioned whether the soil application of common microbiological growth media can influence the plant-microbe interactions and, subsequently, offer protection against V. dahliae. Indeed, the addition of Nutrient Broth (NB) and Potato Dextrose (PD) in non-sterilized soil reduced Verticillium wilt symptoms in eggplants. The addition of NB in sterilized soil did not reduce the disease symptoms compared to controls, however the addition of PD enhanced plant protection against V. dahliae. Following the results of a split root bioassay in eggplants, the possibility that NB and PD triggered the plant defense mechanisms against V. dahliae was excluded, since NB and PD did not reduce wilt symptom. Therefore, PD may be used as an easy food source for V. dahliae, detouring the pathogen from the root system of plants, while NB may affect the soil microbiome by enhancing antagonism in rhizosphere, or antagonistic interaction between V. dahliae and rhizospheric microbiome. Subsequently, several bacterial strains were isolated from the NB-treated rhizosphere and examined for their biocontrol activity against V. dahliae. Among the examined strains, a Pseudomonas putida strain, Z13, significantly reduced Veticillium severity and incidence under greenhouse conditions.

The genus Burkholderia sensu lato is composed of a diverse and metabolically versatile group of bacterial species. One characteristic thought to be unique for the genus Burkholderia is the presence of two forms each (with and without 2-hydroxylation) of the membrane lipids phosphatidylethanolamine (PE) and ornithine lipids (OLs). Here, we show that only Burkholderia sensu stricto strains constitutively form OLs, whereas all other analyzed strains belonging to the Burkholderia sensu lato group constitutively form the two forms of PE, but no OLs. We selected two model bacteria to study the function of OL in Burkholderia sensu lato: (1) Burkholderia cenocepacia wild-type which constitutively forms OLs and its mutant deficient in the formation of OLs and (2) Robbsia andropogonis (formerly Burkholderia andropogonis) which does not form OL constitutively, and a derived strain constitutively forming OLs. Both were characterized under free-living conditions and during pathogenic interactions with their respective hosts. The absence of OLs in B. cenocepacia slightly affected bacterial growth under specific abiotic stress conditions such as high temperature and low pH. B. cenocepacia lacking OLs caused lower mortality in Galleria mellonella larvae while R. andropogonis constitutively forming OLs triggers an increased formation of reactive oxygen species immediately after infection of maize leaves, suggesting that OLs can have an important role during the activation of the innate immune response of eukaryotes.

The study aimed to evaluate the metabolism and resistance to the gastrointestinal tract conditions of Bacillus pumilus UAMX (BP-UAMX) isolated from overweight individuals using genomic tools. Specifically, we assessed its ability to metabolize various carbon sources, its resistance to low pH exposure, and its growth in the presence of bile salts. The genomic and bioinformatic analyses included the prediction of gene and protein metabolic functions, a pan-genome and phylogenomic analysis. BP-UAMX survived at pH 3, while bile salts (0.2–0.3% w/v) increased its growth rate. Moreover, it showed the ability to metabolize simple and complex carbon sources (glucose, starch, carboxymethyl-cellulose, inulin, and tributyrin), showing a differentiated electrophoretic profile. Genome was assembled into a single contig, with a high percentage of genes and proteins associated with the metabolism of amino acids, carbohydrates, and lipids. Antibiotic resistance genes were detected, but only one beta-Lactam resistance protein related to the inhibition of peptidoglycan biosynthesis was identified. The pan-genome of BP-UAMX is still open with phylogenetic similarities with other Bacillus of human origin. Therefore, BP-UAMX seems to be adapted to the intestinal environment, with physiological and genomic analyses demonstrating the ability to metabolize complex carbon sources, the strain has an open pan-genome with continuous evolution and adaptation.

Plants MADS-domain/AGL proteins constitute a large transcription factor (TF) family that controls the development of almost every plant organ. We performed a phylogeny of ( ca . 500) MADS-domain proteins from Arabidopsis and four legume species. We identified clades with Arabidopsis MADS-domain proteins known to participate in root development that grouped legume MADS-proteins with similar high expression in roots and nodules. In this work, we analyzed the role of AGL transcription factors in the common bean ( Phaseolus vulgaris ) – Rhizobium etli N-fixing symbiosis. Sixteen P. vulgaris AGL genes ( PvAGL ), out of 93 family members, are expressed – at different levels – in roots and nodules. From there, we selected the PvAGL gene denominated PvFUL -like for overexpression or silencing in composite plants, with transgenic roots and nodules, that were used for phenotypic analysis upon inoculation with Rhizobium etli . Because of sequence identity in the DNA sequence used for RNAi- FUL- like construct, roots, and nodules expressing this construct -referred to as RNAi_ AGL - showed lower expression of other five PvAGL genes highly expressed in roots/nodules. Contrasting with PvFUL -like overexpressing plants, rhizobia-inoculated plants expressing the RNAi_ AGL silencing construct presented affection in the generation and growth of transgenic roots from composite plants, both under non-inoculated or rhizobia-inoculated condition. Furthermore, the rhizobia-inoculated plants showed decreased rhizobial infection concomitant with the lower expression level of early symbiotic genes and increased number of small, ineffective nodules that indicate an alteration in the autoregulation of the nodulation symbiotic process. We propose that the positive effects of PvAGL TF in the rhizobia symbiotic processes result from its potential interplay with NIN, the master symbiotic TF regulator, that showed a CArG-box consensus DNA sequence recognized for DNA binding of AGL TF and presented an increased or decreased expression level in roots from non-inoculated plants transformed with OE_ FUL or RNAi_ AGL construct, respectively. Our work contributes to defining novel transcriptional regulators for the common bean – rhizobia N-fixing symbiosis, a relevant process for sustainable agriculture.

The genus Weissella is composed of a group of Gram-positive facultative anaerobe bacteria with fermentative metabolism. Strains of this genus have been isolated from various ecological niches, including a wide variety of fermented cereal foods. The present study aimed to determine the relative abundance and fermentation capabilities of Weissella species isolated from pozol, a traditional maya product made of lime-cooked (nixtamalized) fermented maize. We sequenced the V3-V4 regions of 16S rDNA; Weissella was detected early in the fermentation process and reached its highest relative abundance (3.89%) after 3 h of culture. In addition, we evaluated five Weissella strains previously isolated from pozol but reported as non-amylolytic, to define alternative carbon sources such as xylan, xylooligosaccharides, and sucrose. While no growth was observed on birch xylan, growth did occur on xylooligosaccharides and sucrose. Strains WcL17 and WCP-3A were selected for genomic sequencing, as the former shows efficient growth on xylooligosaccharides and the latter displays high glycosyltransferase (GTF) activity. Genomes of both strains were assembled and recorded, with a total of 2.3 Mb in 30 contigs for WcL17 and 2.2 Mb in 45 contigs for WCP-3a. Both strains were taxonomically assigned to Weissella confusa and genomic analyses were performed to evaluate the gene products encoding active carbohydrate enzymes (CAZy). Both strains have the gene content needed to metabolize sucrose, hemicellulose, cellulose, and starch residues, all available in pozol. Our results suggest that the range of secondary enzymatic activity in Weissella confusa strains confer them with wide capabilities to participate in fermentative processes of natural products with heterogeneous carbon sources.

A molecular characterization of a plasmid-born qepA gene in (ESBL)-producing E. coli clinical isolates were performed. An 2.63% (11/418) were qepA positive isolates, of which a 90.0% carried CTX-M-15 (9/11) and SHV-12 (1/11). All isolates showed chromosomal mutations in the gyrA and parC genes. The clonal groups A, B and C were identified and belonged to, respectively, phylogroups A, B1 and D, as well as the sequence types 205, 405 and 617. Several plasmid profiles were determined with incompatibility groups FIA, FIB and FII. The genetic environment of the qepA in plasmid pEC8020 was different from those reported previously. The plasmid sequence included genes conferring resistance to β-lactams ( bla CTX-M-15), macrolides ( mphA ), fluoroquinolones ( qepA1 ), trimethoprim ( dfrB4 ) and sulphonamides ( sul1 ). Likewise, the IncF-pEC8020 plasmid carried several insertion sequences including IS CR 3, IS 6100 and multiple copies of IS 26 . This work contributes to the epidemiology and genetics of plasmid-born qepA genes of ESBL-producing E. coli.

Accurate recognition of the closely related species Klebsiella pneumoniae, Klebsiella quasipneumoniae and Klebsiella variicola by phenotypic, biochemical and automated tests is notoriously unreliable in hospitals' diagnostic laboratories. A comparative genomics approach was conducted for the correct differentiation of the main bacterial species in the K. pneumoniae complex. Analysis of the deduced proteomes of 87 unique genomes of the Klebsiella in public databases, was used for the identification of unique protein family members. This allowed the design of a multiplex-PCR assay for the correct differentiation of these three species from different origins. This system allowed us to determine the prevalence of K. pneumoniae, K. quasipneumoniae and K. variicola among a collection of 552 clinical isolates. Of these, 87.3% (482/552) isolates corresponded to K. pneumoniae, 6.7% (33/552) to K. quasipneumoniae and 5.9% (33/552) to K. variicola. The multiplex-PCR results showed a 100% accuracy for the correct identification of the three species evaluated, which was validated with rpoB phylogenetic sequence analysis.