The nucleoid protein HU positively regulates the expression of type VI secretion systems in Enterobacter cloacae.

Enterobacter cloacae is an emerging pathogen isolated in healthcare-associated infections. A major virulence factor of this bacterium is the type VI secretion system (T6SS). The genome of E. cloacae harbors two T6SS gene clusters (T6SS-1 and T6SS-2), and the functional characterization of both systems showed that these two T6SSs are not expressed under the same conditions. Here, we report that the major histone-like protein HU positively regulates the expression of both T6SSs and, therefore, the function that each T6SS exerts in E. cloacae. Single deletions of the genes encoding the HU subunits (hupA and hupB) decreased mRNA levels of both T6SS. In contrast, the hupA hupB double mutant dramatically affected the T6SS expression, diminishing its transcription. The direct binding of HU to the promoter regions of T6SS-1 and T6SS-2 was confirmed by electrophoretic mobility shift assay. In addition, single and double mutations in the hup genes affected the ability of inter-bacterial killing, biofilm formation, adherence to epithelial cells, and intestinal colonization, but these phenotypes were restored when such mutants were trans-complemented. Our data broaden our understanding of the regulation of HU-mediated T6SS in these pathogenic bacteria. IMPORTANCE: T6SS is a nanomachine that functions as a weapon of bacterial destruction crucial for successful colonization in a specific niche. Enterobacter cloacae expresses two T6SSs required for bacterial competition, adherence, biofilm formation, and intestinal colonization. Expression of T6SS genes in pathogenic bacteria is controlled by multiple regulatory systems, including two-component systems, global regulators, and nucleoid proteins. Here, we reported that the HU nucleoid protein directly activates both T6SSs in E. cloacae, affecting the T6SS-related phenotypes. Our data describe HU as a new regulator involved in the transcriptional regulation of T6SS and its impact on E. cloacae pathogenesis.

The interaction of InvF-RNAP is mediated by the chaperone SicA in Salmonella sp: an in silico prediction.

In this work we carried out an in silico analysis to understand the interaction between InvF-SicA and RNAP in the bacterium Salmonella Typhimurium strain LT2. Structural analysis of InvF allowed the identification of three possible potential cavities for interaction with SicA. This interaction could occur with the structural motif known as tetratricopeptide repeat (TPR) 1 and 2 in the two cavities located in the interface of the InvF and α-CTD of RNAP. Indeed, molecular dynamics simulations showed that SicA stabilizes the Helix-turn-Helix DNA-binding motifs, i.e., maintaining their proper conformation, mainly in the DNA Binding Domain (DBD). Finally, to evaluate the role of amino acids that contribute to protein-protein affinity, an alanine scanning mutagenesis approach, indicated that R177 and R181, located in the DBD motif, caused the greatest changes in binding affinity with α-CTD, suggesting a central role in the stabilization of the complex. However, it seems that the N-terminal region also plays a key role in the protein-protein interaction, especially the amino acid R40, since we observed conformational flexibility in this region allowing it to interact with interface residues. We consider that this analysis opens the possibility to validate experimentally the amino acids involved in protein-protein interactions and explore other regulatory complexes where chaperones are involved.

Interactions between the AraC/XylS-like transcriptional activator InvF of Salmonella Typhimurium, the RNA polymerase alpha subunit and the chaperone SicA.

Salmonella enterica serovar Typhimurium causes gastroenteritis and systemic infections in humans. For this bacterium the expression of a type III secretion system (T3SS) and effector proteins encoded in the Salmonella pathogenicity island-1 (SPI-1), is keystone for the virulence of this bacterium. Expression of these is controlled by a regulatory cascade starting with the transcriptional regulators HilD, HilC and RtsA that induce the expression of HilA, which then activates expression of the regulator InvF, a transcriptional regulator of the AraC/XylS family. InvF needs to interact with the chaperone SicA to activate transcription of SPI-1 genes including sicA, sopB, sptP, sopE, sopE2, and STM1239. InvF very likely acts as a classical activator; however, whether InvF interacts with the RNA polymerase alpha subunit RpoA has not been determined. Results from this study confirm the interaction between InvF with SicA and reveal that both proteins interact with the RNAP alpha subunit. Thus, our study further supports that the InvF/SicA complex acts as a classical activator. Additionally, we showed for the first time an interaction between a chaperone of T3SS effectors (SicA) and the RNAP.

Borrelia turicatae from Ticks in Peridomestic Setting, Camayeca, Mexico.

We conducted surveillance studies in Sinaloa, Mexico, to determine the circulation of tick-borne relapsing fever spirochetes. We collected argasid ticks from a home in the village of Camayeca and isolated spirochetes. Genomic analysis indicated that Borrelia turicatae infection is a threat to those living in resource-limited settings.

Weather and electrical demand and consumption data of a small Mexican community.

The development of novel technologies to mitigate the effects of climate change through Smart Grids requires energy related data. Unfortunately, this type of data is not always available in Mexico, especially from non-large urban areas and at the household level. Therefore, we present a dataset that contains electrical demand and consumption time series of 5 households within a small community in Mexico, at various resolutions, as well as weather data. The electrical demand is given in 15 min resolution, while the electrical consumption is presented in both hourly and daily resolutions. The data is contained within 15 separate .csv files; one for each household's resolution. In turn, the weather data is given in two .csv files (for outdoor and indoor variables, respectively) that together contain 24 meteorological variables measured in a 5 min resolution that is not always consistent. The dataset comprises of two separate folders that contain either the electrical demand and consumption files or the weather files. This dataset could aid in the development of novel smart grid methods and algorithms that might be able to push the energy transition in Mexico and other developing countries forward.

Assessment of chitosan-based adsorbents for glyphosate removal.

Exposure to glyphosate produces various toxic effects, due to this, different methods have been evaluated for its elimination. The objective of this work was to formulate chitosan-based adsorbents and evaluate their efficiency in the removal of glyphosate in vitro. Four films were made by varying the weight ratio of silica/chitosan particles, and four sponges were made by varying the chitosan/chitosan ratio in a reticulated manner. Both adsorbents were characterized based on their porosity, water absorption, glyphosate removal, and reusability. It was found that increasing the porosity in both films and sponges resulted in an increase in the adsorption efficiency of glyphosate. The adsorption process exhibited a better fit in both adsorbents to the pseudo-second-order model. The adsorption of glyphosate to the films fit better with the Langmuir model, demonstrating that the process occurs in the form of a monolayer. In the case of sponges, the adsorption of glyphosate fit better with the Freundlich model, indicating that the process takes place in a multilayer form. Finally, when the reusability was evaluated, the adsorbents showed a loss of effectiveness. However, they still proved to be an efficient alternative for the removal of glyphosate in water, providing a cost-effective and environmentally friendly solution.

Mathematical modeling for operative improvement of the decoloration of Acid Red 27 by a novel microbial consortium of Trametes versicolor and Pseudomonas putida: A multivariate sensitivity analysis.

In this work, it is presented a first approach of a mathematical and kinetic analysis for improving the decoloration and further degradation process of an azo dye named acid red 27 (AR27), by means of a novel microbial consortium formed by the fungus Trametes versicolor and the bacterium Pseudomonas putida. A multivariate analysis was carried out by simulating scenarios with different operating conditions and developing a specific mathematical model based on kinetic equations describing all stages of the biological process, from microbial growth and substrate consuming to decoloration and degradation of intermediate compounds. Additionally, a sensitivity analysis was performed by using a factorial design and the Response Surface Method (RSM), for determining individual and interactive effects of variables like, initial glucose concentration, initial dye concentration and the moment in time for bacterial inoculation, on response variables assessed in terms of the minimum time for: full decoloration of AR27 (R1 = 2.375 days); maximum production of aromatic metabolites (R2 = 1.575 days); and full depletion of aromatic metabolites (R3 = 12.9 days). Using RSM the following conditions improved the biological process, being: an initial glucose concentration of 20 g l-1, an initial AR27 concentration of 0.2 g l-1 and an inoculation moment in time of P. putida at day 1. The mathematical model is a feasible tool for describing AR27 decoloration and its further degradation by the microbial consortium of T. versicolor and P. putida, this model will also work as a mathematical basis for designing novel bio-reaction systems than can operate with the same principle of the described consortium.

New records of Ornithodoros turicata (Ixodida: Argasidae) in rural and urban sites in the Mexican states of Aguascalientes and Zacatecas indicate the potential for tick-borne relapsing fever.

Soft ticks from the Ornithodoros genus are vectors of relapsing fever (RF) spirochetes around the world. In Mexico, they were originally described in the 19th century. However, few recent surveillance studies have been conducted in Mexico, and regions where RF spirochetes circulate remain vague. Here, the presence of soft ticks in populated areas was assessed in two sites from the Mexican states of Aguascalientes and Zacatecas. Argasidae ticks were collected, identified by morphology and mitochondrial 16S rDNA gene sequencing, and tested for RF borreliae. The specimens in both sites were identified as Ornithodoros turicata but no RF spirochetes were detected. These findings emphasize the need to update the distribution of these ticks in multiple regions of Mexico and to determine the circulation of RF borreliosis in humans and domestic animals.

Detection of Rickettsia amblyommatis and Rickettsia bellii in ticks collected from pet dogs in peri-urban and rural areas in Yucatan, Mexico.

Rickettsia species are bacteria that may cause multiple diseases in animals and humans, via transmission through multiple arthropod vectors. Routine surveillance of Rickettsia spp. within vectors is critical to determine their presence and risk to mammalian hosts within human populations. Therefore, to better characterize the circulating Rickettsia species in an understudied region we targeted pet dogs to survey. Ticks were collected from pet dogs in three populations of the Yucatan where we tested for the presence of Rickettsia spp. by PCR in metagenomic DNA. In these ticks removed from pet dogs we detected Rickettsia amblyommatis and Rickettsia bellii in Amblyomma auriculatum, Amblyomma ovale and Amblyomma mixtum ticks obtained in a rural community in the Mexican state of Yucatan. This is the first report detecting both species for this state in Mexico, underpinning the importance of more routine surveillance.

Case Report: Exposure to Relapsing Fever Group Borreliae in Patients with Undifferentiated Febrile Illness in Mexico.

Relapsing fever (RF) borreliosis is a neglected disease in Mexico. A retrospective serological survey using diagnostic antigens GlpQ and BipA from Borrelia turicatae was performed to evaluate human exposure to RF borreliae. Seventy serum samples were used from a cohort of patients with undifferentiated febrile illness in Mexico. Four samples were positive to GlpQ and three to BipA. Results indicate that RF borreliae continue to circulate in regions of Mexico and pose a risk to human health.

Distribution of CRISPR-Cas systems in the Burkholderiaceae family and its biological implications.

CRISPR-Cas systems are composed of repeated sequences separated by non-repeated sequences that are near genes coding for Cas proteins, which are involved in the function of these systems. Their function has been mostly related to "genetic immunity" against foreign genetic material, among other roles. Interest in them increased after their use in genetic manipulation was uncovered and surveys to find and classify them have been done in several bacterial groups. To determine the presence of these genetic elements in the Burkholderiaceae family members, a bioinformatic approach was followed. Attention in this family comes as it is formed by a great diversity of microorganisms that include opportunistic and true pathogens, and symbiotic and saprophytic organisms, among others. Results show that, in contrast to other bacterial groups, only 8.4% of family members harbor complete CRISPR-Cas systems and the rest either do not have one or have remains or sections of one. Analyses of the spacer sequences indicated that most of them have identity to sections of the same genomes they were found, while a few had identities with either plasmids or phages. The genus with the higher proportion of self-directed spacers is Ralstonia, and their possible roles are discussed. Most of the systems (60%) belong to the class I subtype I-E and a few to subtypes I-C (13.3%), I-F (18.3%), II-C (5%), IV-A (1.7%) and V-C (1.7%). To the best of our knowledge, this is the first study to uncover the CRISPR-Cas system for the whole Burkholderiaceae family.

Life History Data of Dipetalogaster maxima (Hemiptera: Reduviidae).

Dipetalogaster maxima (Uhler) is a triatomine species that has been found to be infected by Trypanosoma cruzi Chagas in the habitats of the most important tourist areas of Mexico. Its behavior and vectorial capacity have been scarcely studied, although such information is necessary to reliably estimate the importance of this species as a vector of T. cruzi in its distribution area. This study reports biological parameters related to the vectorial capacity of D. maxima. In particular, the egg-to-adult development time, number of blood meals required to molt, accumulative mortality, time to beginning of feeding, feeding and defecation times, fecundity, and fertility were examined. D. maxima took a median of 211 d to develop from egg to adult, requiring 11 meals in total. Almost two-thirds (63%) of specimens died during the cycle. The time to beginning of feeding was 1 min in all instars. Feeding times varied from 14 to 27 min. Most nymphs (except first-instar) defecated when feeding or immediately thereafter. A mean of 0.7 eggs/♀/day was recorded, with an eclosion rate of 27.3%. Five of the eight studied parameters (mainly defecation delay) suggest the remarkable potential vectorial capacity of D. maxima, so it is necessary to maintain permanent surveillance of domiciliary populations of D. maxima, because they may be infected with T. cruzi.

A New Record of the Introduced Species Triatoma infestans (Hemiptera: Reduviidae) in Mexico.

Here, we report a new record of Triatoma infestans (Klug) in Mexico after 50 years and provide a brief description of the discovery area. Fifty-nine specimens (71.2% adults) of the introduced species were collected from the peridomestic areas of a single house in the port of Manzanillo in the state of Colima, Mexico. Thirty-one specimens (52.5%) were collected from the exterior walls of the house and were apparently attracted to light. The other specimens (47.5%) were associated with chickens. No specimen was infected with Trypanosoma cruzi Chagas, the causative agent of Chagas disease, possibly because they were feeding on chickens. We speculate that the introduced species travelled from South America to Mexico via seed shipment in a twenty-foot equivalent unit (TEU) maritime container. Because Mexican phytosanitary regulations demand only the cargo to be inspected, the triatomines could have escaped notice during inspection. Subsequently, as the cargo was unloaded and the TEU was stored, the triatomines likely flew to and invaded the nearby residential areas. The rediscovery of this domestic vector of T. cruzi in Mexico warrants further investigation owing to the potential risk of transmission to the inhabitants of the study area.

Burkholderia orbicola sp. nov., a novel species within the Burkholderia cepacia complex.

Genome analysis of strains placed in the NCBI genome database as Burkholderia cenocepacia defined nine genomic species groups. The largest group (259 strains) corresponds to B. cenocepacia and the second largest group (58 strains) was identified as "Burkholderia servocepacia", a Burkholderia species classification which has not been validly published. The publication of "B. servocepacia" did not comply with Rule 27 and Recommendation 30 from the International Code of Nomenclature of Prokaryotes (ICNP) and have errors in the type strain name and the protologue describing the novel species. Here, we correct the position of this species by showing essential information that meets the criteria defined by ICNP. After additional analysis complying with taxonomic criteria, we propose that the invalid "B. servocepacia" be renamed as Burkholderia orbicola sp. nov. The original study proposing "B. servocepacia" was misleading, because this name derives from the Latin "servo" meaning "to protect/watch over", and the authors proposed this based on the beneficial biocontrol properties of several strains within the group. However, it is clear that "B. servocepacia" isolates are capable of opportunistic infection, and the proposed name Burkholderia orbicola sp. nov. takes into account these diverse phenotypic traits. The type strain is TAtl-371 T (= LMG 30279 T = CM-CNRG 715 T).

Biological Parameters of Two Triatoma protracta Subspecies (Hemiptera: Reduviidae).

In recent years, concerns about Chagas disease in the United States have increased. Triatomine bug (Hemiptera: Reduviidae) populations are the vectors of the parasite Trypanosoma cruzi Chagas (Trypanosomatida: Trypanosomatidae), which causes Chagas disease, although the route of transmission is considered inefficient in United States. However, more studies on triatomine feeding and defecation behavior are needed. In this study, six related biological parameters from two populations of Triatoma protracta protracta (Uhler) and T. p. woodi (Uhler) from Mexican locations near the U.S. border were evaluated. The four population life cycles were less than 6 mo (161-171 d), with 9-10 blood meals needed to molt. Mortality rates were similar (31-38%) among the four populations. Triatoma p. woodi from Hidalgo, Coahuila was the most aggressive one. Feeding times were over 10 min, increasing with instar in all populations. Defecation behaviors varied among populations. High percentages of male and female fourth- and fifth-instar nymphs of T. p. protracta from Imuris and both populations of T. p. woodi defecated immediately after or <1 min of feeding. Lower percentages were observed for T. p. protracta from Jacumé. Because most parameters were similar among the four populations, independent of their subspecies and their geographic origin, we considered that T. p. protracta and T. p. woodi are efficient vectors of T. cruzi. In contrast, defecation patterns were noticeably different among some of the four triatomine populations studied. Our results highlight the importance of studying the biological parameters of local triatomine populations. They also contribute to increasing the knowledge of North American triatomine behavior and defecation patterns.

Long-chain fatty acids alter transcription of Helicobacter pylori virulence and regulatory genes.

Infection with Helicobacter pylori is one of the most important risk factors for developing gastric cancer (GC). The type IV secretion system (T4SS) encoded in the cag pathogenicity island is the main virulence factor of H. pylori associated with GC. Additionally, other virulence factors have been shown to play a role in the H. pylori virulence, such as vacuolizing cytotoxin (VacA), urease, flagella, and adhesins. Long-chain fatty acids (LCFAs) are signaling molecules that affect the transcription of virulence genes in several pathogenic bacteria such as Salmonella enterica, Vibrio cholerae, Pseudomonas aeruginosa and Mycobacterium tuberculosis. However, the effect of LCFAs on the transcription of H. pylori virulence and regulatory genes remains unknown. Here we analyzed whether the transcription of virulence genes that encode T4SS and cellular envelope components, flagellins, adhesins, toxins, urease, as well as the transcription of different regulatory genes of the H. pylori strain 26695, are altered by the presence of five distinct LCFAs: palmitic, stearic, oleic, linoleic, and linolenic acids. Palmitic and oleic acids up-regulated the transcription of most of the virulence genes tested, including cagL, cagM, flaB, sabA, mraY and vacA, as well as that of the genes encoding the transcriptional regulators NikR, Fur, CheY, ArsR, FlgR, HspR, HsrA, Hup, and CrdR. In contrast, the other LCFAs differentially affected the transcription of the virulence and regulatory genes assessed. Our data show that LCFAs can act as signaling molecules that control the transcription of the H. pylori virulome.

Point mutations in Candida glabrata 3-hydroxy-3-methylglutaryl-coenzyme A reductase (CgHMGR) decrease enzymatic activity and substrate/inhibitor affinity.

3-Hydroxy-3-methylglutaryl-coenzyme A reductase (HMGR) is a crucial enzyme in the ergosterol biosynthesis pathway. The aim of this study was to obtain, purify, characterize, and overexpress five point mutations in highly conserved regions of the catalytic domain of Candida glabrata HMGR (CgHMGR) to explore the function of key amino acid residues in enzymatic activity. Glutamic acid (Glu) was substituted by glutamine in the E680Q mutant (at the dimerization site), Glu by glutamine in E711Q (at the substrate binding site), aspartic acid by alanine in D805A, and methionine by arginine in M807R (the latter two at the cofactor binding site). A double mutation, E680Q-M807R, was included. Regarding recombinant and wild-type CgHMGR, in vitro enzymatic activity was significantly lower for the former, as was the in silico binding energy of simvastatin, alpha-asarone and the HMG-CoA substrate. E711Q displayed the lowest enzymatic activity and binding energy, suggesting the importance of Glu711 (in the substrate binding site). The double mutant CgHMGR E680Q-M807R exhibited the second lowest enzymatic activity. Based on the values of the kinetic parameters KM and Vmax, the mutated amino acids appear to participate in binding. The current findings provide insights into the role of residues in the catalytic site of CgHMGR.

Biological Parameters of Three Populations of Triatoma dimidiata s. s. (Hemiptera: Reduviidae) From Western Mexico.

Chagas disease is a very important vector-borne disease in México, and Triatoma dimidiata sensu stricto (Latreille) is one of the most important vectors of Trypanosoma cruzi Chagas, mainly in southern and central states. In the Pacific Coast states with the highest prevalence of human T. cruzi infection, T. dimidiata s. s. is considered as a secondary vector. However, the vectorial capacity of those populations has not been studied. Therefore, the vector characteristics of three populations of T. dimidiata s. s. in western México were evaluated in this study. The populations were maintained in the laboratory at 27 ± 1°C and 75% ± 5% RH with a 12:12 h (light:dark) regime, fed on rabbits in a fortnight basis. The development times were short (172-238 d), and the number of bloodmeals to molt was low (11). Mortality was moderate (36-45%), the onset of feeding was relatively rapid (0.5-1.7 min), and feedings were extended (>15 min). More than 40% of individuals in most instars defecated in one of three categories: <1 min when feeding (5-37.9%), immediately after feeding (9-28.6%), or in <1 min post feeding (7-25.8%). The median number of laid eggs was high (over 2.5) in the three populations, as were the egg eclosion rates (>86%). Thus, the T. dimidiata s. s. in the three populations are potentially efficient vectors of T. cruzi and could contribute to the high prevalence of infection in human populations in western México.

Burkholderia species in human infections in Mexico: Identification of B. cepacia, B. contaminans, B. multivorans, B. vietnamiensis,B. pseudomallei and a new Burkholderia species.

BACKGROUND: Burkholderia sensu stricto is comprised mainly of opportunistic pathogens. This group is widely distributed in the environment but is especially important in clinical settings. In Mexico, few species have been correctly identified among patients, most often B. cepacia is described. METHODOLOGY/PRINCIPAL FINDINGS: In this study, approximately 90 strains identified as B. cepacia with the VITEK2 system were isolated from two medical centers in Mexico City and analyzed by MLSA, BOX-PCR and genome analysis. The initial identification of B. cepacia was confirmed for many strains, but B. contaminans, B. multivorans and B. vietnamiensis were also identified among clinical strains for the first time in hospitals in Mexico. Additionally, the presence of B. pseudomallei was confirmed, and a novel species within the B. cepacia complex was documented. Several strains misidentified as B. cepacia actually belong to the genera Pseudomonas, Stenotrophomonas and Providencia. CONCLUSIONS/SIGNIFICANCE: The presence of different Burkholderia species in Mexico was confirmed. Correct identification of Burkholderia species is important to provide accurate treatment for immunosuppressed patients.

An update of the unceasingly growing and diverse AraC/XylS family of transcriptional activators.

Transcriptional factors play an important role in gene regulation in all organisms, especially in Bacteria. Here special emphasis is placed in the AraC/XylS family of transcriptional regulators. This is one of the most abundant as many predicted members have been identified and more members are added because more bacterial genomes are sequenced. Given the way more experimental evidence has mounded in the past decades, we decided to update the information about this captivating family of proteins. Using bioinformatics tools on all the data available for experimentally characterized members of this family, we found that many members that display a similar functional classification can be clustered together and in some cases they have a similar regulatory scheme. A proposal for grouping these proteins is also discussed. Additionally, an analysis of surveyed proteins in bacterial genomes is presented. Altogether, the current review presents a panoramic view into this family and we hope it helps to stimulate future research in the field.