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The genus Rhytidhysteron is characterised by forming navicular to apothecial hysterothecia, exposing the green, yellow, orange, red, vinaceous or black colours of the hymenium which generally releases pigments in the presence of KOH. The exciple is smooth or striated, the asci bitunicate and ascospores have 1–5 transverse septa. To date, twenty-six Rhytidhysteron species have been described from the Tropics. The present study aims to describe three new species in the Neotropics of Mexico based on molecular methods and morphological features. Illustrations and a taxonomic key are provided for all known species of this genus. Rhytidhysteron cozumelense from the Isla Cozumel Biosphere Reserve, R. esperanzae from the Sierra Juárez, Oaxaca and R. mesophilum from the Sierra Madre Oriental, Hidalgo are described as new species. With the present study, the number of species of Rhytidhysteron known from Mexico is now increased to eight.

Integrons are prokaryotic genetic elements known to carry and exchange antibiotic resistance gene cassettes through a site-specific recombinase called integrase. In this work, 107 Aeromonas isolates from environmental origin, including fish, water, and sediments, were investigated for the presence of integrons. Using specific primers for Class 1, 2 and 3 integrases, only Class 1 and Class 2 integrons were detected. Detection of Class 2 integrases and their associated variable regions required two rounds of polymerase chain reaction (PCR). Sequencing of the intI2 amplicons confirmed them as integrase-derived products. Class 1 integrons were detected in 26 out of 107 isolates. PCR amplification of the variable regions associated to these integrons revealed an outstanding homogeneity, 25 of them having variable regions with an identical dfrA12-orfF-aadA2 cassette array and one integron carrying only the dfrA16 cassette. To assess clone diversity, chromosomal DNA from isolates was subjected to enterobacterial repetitive intergenic consensus-PCR (ERIC-PCR), which discarded clonality in all instances. Class 2 integrons were surprisingly more prevalent than Class1 integrons, being detected in 60 out of 107 isolates. Forty-six of them showed a unique ERIC profile, while the remaining 14 strains displayed profiles that could be grouped in five different patterns. Cassette arrangements of all Class 2 variable regions were those described as the most prevalent (dfrA1-sat2-aadA1). A rather startling result of this work is the sensitivity to trimethoprim, streptomycin, and streptothricin of most strains, despite the presence of the cognate resistance genes. To know the integron distribution in environmental Aeromonas species, a phylogenetic reconstruction was done using rpoD/gyrB or rpoD/gyrA gene sequences. Isolates bearing these elements corresponded to Aeromonas hydrophila, Aeromonas veronii, Aeromonas salmonicida, Aeromonas dhakensis, Aeromonas sanarellii, Aeromonas taiwanensis, Aeromonas media, Aeromonas caviae, Aeromonas jandaei, and Aeromonas sp. This work revealed an unusual high incidence of Class 2 integrons and a low variability of cassette arrangements in environmental Aeromonas species.

Plasmids are autonomous genetic elements ubiquitously present in bacteria. In addition to containing genetic determinants responsible for their replication and stability, some plasmids may carry genes that help bacteria adapt to different environments, while others without a known function are classified as cryptic. In this work we identified and characterized plasmids from a collection of mesophilic Aeromonas spp. (N = 90) isolated from water, sediments and fish. A total of 15 small plasmids ranging from 2287 to 10,558 bp, with an incidence of 16.7% (15/90) was found. Plasmids were detected in A. hydrophila (6), A. veronii (4), A. taiwanensis (2), A. jandaei (1), A. media (1) and Aeromonas sp. (1). There were no large or megaplasmids in the strains studied in this work. Analysis of coding sequences identified proteins associated to replication, mobilization, antibiotic resistance, virulence and stability. A considerable number of hypothetical proteins with unknown functions were also found. Some strains shared identical plasmid profiles, however, only two of them were clones. Small plasmids could be acting as a gene repositories as suggested by the presence of a gene encoding for a putative zonula occludens toxin (Zot) that causes diarrhea and the qnrB gene involved in quinolone resistance harbored in plasmids pAerXII and pAerXIII respectively.

Resumen:Antecedentes y Objetivos: El género Daldinia, con 48 especies a nivel mundial, se caracteriza por presentar peritecios inmersos en estromas globosos a clavados de colores cobrizos, violáceos, vináceos, marrones a negros, liberando pigmentos de colores ocráceos, púrpuras, verdosos y grisáceos en KOH. Está caracterizado por ascas cilíndricas estipitadas con poro apical amiloide, y ascosporas elipsoidales marrones lisas u ornamentadas. El estado asexual fue descrito en el género Nodulisporium. Actualmente, se sabe que el grupo es principalmente endófito. El objetivo del presente estudio fue realizar una revisión taxonómica del género Daldinia en México, basada en caracteres macro y micromorfológicos y la observación de las esporas al MEB para identificar a las especies.Métodos: Los ejemplares fueron estudiados, descritos y determinados de acuerdo con las técnicas tradicionales en micología. Adicionalmente, las esporas se observaron al microscopio electrónico de barrido. El material está depositado en la colección de hongos del herbario ENCB.Resultados clave: Se determinaron 14 especies de Daldinia en México; de éstas, D. decipiens, D. cuprea, D. petriniae y D. rehmii se citan por primera vez para el país. De acuerdo con el hábitat, las especies de este género se asociaron principalmente al bosque tropical caducifolio. Daldinia childiae, D. eschscholtzii y D. vernicosa son las especies con mejor distribución en el territorio nacional.Conclusiones: El género Daldinia se encuentra bien representado en México, siendo uno de los mejor estudiados de la familia Hypoxylaceae. El género tiene una distribución principalmente tropical, con más de 80% de las especies descritas. En regiones templadas su distribución es restringida y manifiesta una mayor especificidad por el hospedero, principalmente Alnus y Quercus. El estado de Oaxaca fue la entidad con mayor riqueza de especies.

Integrons are prokaryotic genetic elements known to carry and exchange antibiotic resistance gene cassettes through a site-specific recombinase called integrase. In this work, 107

Staphylococcus aureus is a commensal bacterium in humans and animals able to adapt to multiple environments. The aim of this study was to compare the genetic diversity and virulence profiles of strains of S. aureus isolated from food (29 strains), humans (43 strains), and animals (8 strains). 80 lipase-producing strains belonging to a biobank of 360 isolates, identified phenotypically as S. aureus, were selected. Confirmation of the species was made by amplifying the spA gene and 80% (64/80) of the strains were confirmed within this species. The virulence profile of each of the isolates was determined by PCR. The seA gene coding for enterotoxin A was found in 53.1% of the strains, the saK gene, which codes for Staphylokinase, was amplified in 57.8% of the strains, and, finally, the hlB gene coding for β-Hemolysin was amplified in 17.2%. The profile of antimicrobial resistance was determined by the Kirby Bauer method showing that the strains from food presented greater resistance to erythromycin (40.7%) and ciprofloxacin (18.5%) while in strains isolated from humans were to erythromycin (48.4%) and clindamycin (21.2%). Also, in strains from animals, a high resistance to erythromycin was observed (75%). The frequency of MRSA was 12.5% due to the presence of the mec gene and resistance to cefoxitin. Of the total strains, 68.7% were typed by PCR-RFLP of the coa gene using the AluI enzyme; derived from this restriction, 17 profiles were generated. Profile 4 (490 bp, 300 bp) was the most frequent, containing a higher number of strains with a higher number of virulence factors and antimicrobial resistance, which is associated with greater adaptation to different environments. In this study, a wide genetic diversity of strains of S. aureus from different foods, humans, and animals was found. This demonstrates evolution, genetic versatility, and, therefore, the adaptation of this microorganism in different environments.

In 1995, Pseudomonas sp. ADP, capable of metabolizing atrazine, was isolated from contaminated soil. Genes responsible for atrazine mineralization were found scattered in the 108.8 kb pADP-1 plasmid carried by this strain, some of them flanked by insertion sequences rendering them unstable. The goal of this work was to construct a transcriptional unit containing the atz operon in an easy to transfer manner, to be introduced and inherited stably by Gram-negative bacteria. atz genes were PCR amplified, joined into an operon and inserted onto the mobilizable plasmid pBAMD1–2. Primers were designed to add efficient transcription and translation signals. Plasmid bearing the atz operon was transferred to different Gram-negative strains by conjugation, which resulted in Tn5 transposase-mediated chromosomal insertion of the atz operon. To test the operon activity, atrazine degradation by transposants was assessed both colorimetrically and by high-performance liquid chromatography (HPLC). Transposants mineralized atrazine more efficiently than wild-type Pseudomonas sp. ADP and did not accumulate cyanuric acid. Atrazine degradation was not repressed by simple nitrogen sources. Genes conferring atrazine-mineralizing capacities were stable and had little or null effect on the fitness of different transposants. Introduction of catabolic operons in a stable fashion could be used to develop bacteria with better degrading capabilities useful in bioremediation.

Major perturbations in soil and water quality are factors that can negatively impact human health. In soil environments of urban areas, changes in antibiotic-resistance profiles may represent an increased risk of exposure to antibiotic-resistant bacteria via oral, dermal, or inhalation routes. We studied the perturbation of antibiotic-resistance profiles and microbial communities in soils following a major flooding event in Houston, Texas, caused by Hurricane Harvey. The main objective of this study was to examine the presence of targeted antibiotic-resistance genes and changes in the diversity of microbial communities in soils a short time (3–5 months) and a long time (18 months) after the catastrophic flooding event. Using polymerase chain reaction, we surveyed fourteen antibiotic-resistance elements: intI1, intI2, sul1, sul2, tet(A) to (E), tet(M), tet(O), tet(W), tet(X), and blaCMY-2. The number of antibiotic-resistance genes detected were higher in short-time samples compared to samples taken a long time after flooding. From all the genes surveyed, only tet(E), blaCMY-2, and intI1 were prevalent in short-time samples but not observed in long-time samples; thus, we propose these genes as indicators of exogenous antibiotic resistance in the soils. Sequencing of the V3-V4 region of the bacterial 16S rRNA gene was used to find that flooding may have affected bacterial community diversity, enhanced differences among bacterial lineages profiles, and affected the relative abundance of Actinobacteria, Verrucomicrobia, and Gemmatimonadetes. A major conclusion of this study is that antibiotic resistance profiles of soil bacteria are impacted by urban flooding events such that they may pose an enhanced risk of exposure for up to three to five months following the hurricane. The occurrence of targeted antibiotic-resistance elements decreased eighteen months after the hurricane indicating a reduction of the risk of exposure long time after Harvey.

Integrons are non-mobile bacterial genetic elements that carry different cassettes conferring antibiotic resistance. Cassettes can excise or integrate by action of an integron-encoded integrase, enabling bacteria to face environmental challenges. In this work, the functionality and dynamics of two integrons carrying the same cassette arrangement (dfrA12–orfF–aadA2), but located on plasmid or chromosome in two different strains were studied. In order to demonstrate the functionality of the Class 1 integrase, circular cassette integration intermediaries were PCR amplified by PCR using extrachromosomal DNA extracted from bacteria grown in the presence or absence of cassette-encoded antibiotics. Circular aadA2 and dfrA12–orfF–aadA2 cassettes were detected in cultures grown either in the presence or absence of antibiotics in both strains. No dfrA12–orfF circular intermediates could be detected under any culture conditions. These results show that both integrons are functional. However, these elements show different dynamics and functionality since the presence of streptomycin led to detectable gene rearrangements in the variable region only in the strain with the plasmid-born integron. In addition, complete integration products were demonstrated using a receptor molecule carrying an empty integron. In this case, integration products were observed in both strains even in the absence of antibiotics, but they were more evident in the strain with the plasmid-located integron when streptomycin was present in the culture medium. This suggests that integrons in the two strains respond differently to streptomycin even though DNA sequences upstream the intI1 gene, including the lexA boxes of both integrons are identical.

Integrons are non-mobile bacterial genetic elements that carry different cassettes conferring antibiotic resistance. Cassettes can excise or integrate by action of an integron-encoded integrase, enabling bacteria to face environmental challenges. In this work, the functionality and dynamics of two integrons carrying the same cassette arrangement (dfrA12–orfF–aadA2), but located on plasmid or chromosome in two different strains were studied. In order to demonstrate the functionality of the Class 1 integrase, circular cassette integration intermediaries were PCR amplified by PCR using extrachromosomal DNA extracted from bacteria grown in the presence or absence of cassette-encoded antibiotics. Circular aadA2 and dfrA12–orfF–aadA2 cassettes were detected in cultures grown either in the presence or absence of antibiotics in both strains. No dfrA12–orfF circular intermediates could be detected under any culture conditions. These results show that both integrons are functional. However, these elements show different dynamics and functionality since the presence of streptomycin led to detectable gene rearrangements in the variable region only in the strain with the plasmid-born integron. In addition, complete integration products were demonstrated using a receptor molecule carrying an empty integron. In this case, integration products were observed in both strains even in the absence of antibiotics, but they were more evident in the strain with the plasmid-located integron when streptomycin was present in the culture medium. This suggests that integrons in the two strains respond differently to streptomycin even though DNA sequences upstream the intI1 gene, including the lexA boxes of both integrons are identical.

We determined the presence of class 1 integrons related to the acquisition of resistance to antimicrobials in Aeromonas spp. isolated from individuals with diarrhea. Species were identified as A. caviae, A. hydrophila, A. veronii and A. media using PCR-RFLP of the 16S rDNA. Selected isolates were further characterized by ERIC-PCR. Resistance to chloramphenicol, aztreonam, tetracycline, trimethoprim/sulfamethoxazole, nalidixic acid and streptomycin, among others, was determined using the Kirby-Bauer method. Integrons were detected by PCR amplification of the 5' conserved, variable, and 3' conserved regions. Sequencing of the variable regions revealed class 1 integrons with cassettes encoding resistance to trimethoprim (dfrA12, dfrA15, dfrB4), streptomycin/spectinomycin (aadA2, aadA1), oxacillin (oxa2) and chloramphenicol (catB3, cmlA4). Others had an open reading frame (orfD) or no insert at all. To our knowledge, this is the first description of the occurrence of genes cmlA4 and dfrA15 in Aeromonas class 1 integrons. Not all the integron-linked cassettes conferred their associated resistances, which suggests the inactivity of some cassettes. Most integrons were chromosomally located. The presence of class 1 integrons similar to those found in a wide variety of bacterial genera from different origins, including environmental and fish-borne Aeromonas, confirms the stability and horizontal transfer of these genetic elements.

Mercury-resistant Aeromonas strains isolated from diarrhea were studied. Resistance occurs via mercuric ion reduction but merA and merR genes were only detected in some strains using PCR and dot hybridization. Results indicate a high variability in mer operons in Aeromonas. To our knowledge, this is the first report of mercury-resistant clinical Aeromonas strains.