Pseudomonas aeruginosa Isolates from Water Samples of the Gulf of Mexico Show Similar Virulence Properties but Different Antibiotic Susceptibility Profiles than Clinical Isolates.

Pseudomonas aeruginosa is an opportunistic pathogen found in a wide variety of environments, including soil, water, and habitats associated with animals, humans, and plants. From a One Health perspective, which recognizes the interconnectedness of human, animal, and environmental health, it is important to study the virulence characteristics and antibiotic susceptibility of environmental bacteria. In this study, we compared the virulence properties and the antibiotic resistance profiles of seven isolates collected from the Gulf of Mexico with those of seven clinical strains of P. aeruginosa. Our results indicate that the marine and clinical isolates tested exhibit similar virulence properties; they expressed different virulence factors and were able to kill Galleria mellonella larvae, an animal model commonly used to analyze the pathogenicity of many bacteria, including P. aeruginosa. In contrast, the clinical strains showed higher antibiotic resistance than the marine isolates. Consistently, the clinical strains exhibited a higher prevalence of class 1 integron, an indicator of anthropogenic impact, compared with the marine isolates. Thus, our results indicate that the P. aeruginosa marine strains analyzed in this study, isolated from the Gulf of Mexico, have similar virulence properties, but lower antibiotic resistance, than those from hospitals.

The alternative sigma factor RpoS regulates Pseudomonas aeruginosa quorum sensing response by repressing the pqsABCDE operon and activating vfr.

Pseudomonas aeruginosa is an important opportunistic pathogen. Several of its virulence-related processes, including the synthesis of pyocyanin (PYO) and biofilm formation, are controlled by quorum sensing (QS). It has been shown that the alternative sigma factor RpoS regulates QS through the reduction of lasR and rhlR transcription (encoding QS regulators). However, paradoxically, the absence of RpoS increases PYO production and biofilm development (that are RhlR dependent) by unknown mechanisms. Here, we show that RpoS represses pqsE transcription, which impacts the stability and activity of RhlR. In the absence of RpoS, rhlR transcript levels are reduced but not the RhlR protein concentration, presumably by its stabilization by PqsE, whose expression is increased. We also report that PYO synthesis and the expression of pqsE and phzA1B1C1D1E1F1G1 operon exhibit the same pattern at different RpoS concentrations, suggesting that the RpoS-dependent PYO production is due to its ability to modify PqsE concentration, which in turn modulates the activation of the phzA1 promoter by RhlR. Finally, we demonstrate that RpoS favors the expression of Vfr, which activates the transcription of lasR and rhlR. Our study contributes to the understanding of how RpoS modulates the QS response in P. aeruginosa, exerting both negative and positive regulation.

Marginal versus Segmental Mandibulectomy in the Treatment of Oral Cavity Cancer: A Systematic Review and Meta-analysis.

Introduction Oral cavity squamous cell carcinoma (OCSCC) is the most common malignancy in the oral cavity. Two types of mandibular resections have been described: the segmental mandibulectomy and the marginal mandibulectomy. Both may have a different impact over the quality of life, oncological prognosis, and functional or aesthetic result. Objectives The aim of this study was to systematically explore the literature to determine the survival outcomes and disease control rates in patients who underwent segmental or marginal mandibulectomy for OCSCC with histological evidence of cortical and medullary bone invasion. Data Synthesis This review involved a systematic search of the electronic databases MEDLINE/PUBMED, Google Scholar, Ovid Medline, Embase, and Scopus including articles from 1985 to 2019. Fifteen articles were included for qualitative analysis and 11 articles were considered for meta-analysis calculations. All of them correspond to retrospective cohort studies. Conclusion This systematic review reveals the low-level evidence regarding the impact over local control or survival according to the type of mandibulectomy. Our results need to be considered with precaution according to the limited evidence available. We just found difference regarding the 5-year disease-free survival, and a tendency in favor of segmental mandibulectomy was confirmed when medullary invasion was evident.

To cheat or not to cheat: cheatable and non-cheatable virulence factors in Pseudomonas aeruginosa.

Important bacterial pathogens such as Pseudomonas aeruginosa produce several exoproducts such as siderophores, degradative enzymes, biosurfactants, and exopolysaccharides that are used extracellularly, benefiting all members of the population, hence being public goods. Since the production of public goods is a cooperative trait, it is in principle susceptible to cheating by individuals in the population who do not invest in their production, but use their benefits, hence increasing their fitness at the expense of the cooperators' fitness. Among the most studied virulence factors susceptible to cheating are siderophores and exoproteases, with several studies in vitro and some in animal infection models. In addition to these two well-known examples, cheating with other virulence factors such as exopolysaccharides, biosurfactants, eDNA production, secretion systems, and biofilm formation has also been studied. In this review, we discuss the evidence of the susceptibility of each of those virulence factors to cheating, as well as the mechanisms that counteract this behavior and the possible consequences for bacterial virulence.

Exoprotease exploitation and social cheating in a Pseudomonas aeruginosa environmental lysogenic strain with a noncanonical quorum sensing system.

Social cheating is the exploitation of public goods that are costly metabolites, like exoproteases. Exoprotease exploitation in Pseudomonas aeruginosa has been studied in reference strains. Experimental evolution with reference strains during continuous growth in casein has demonstrated that nonexoprotease producers that are lasR mutants are selected while they behave as social cheaters. However, noncanonical quorum-sensing systems exist in P. aeruginosa strains, which are diverse. In this work, the exploitation of exoproteases in the environmental strain ID4365 was evaluated; ID4365 has a nonsense mutation that precludes expression of LasR. ID4365 produces exoproteases under the control of RhlR, and harbors an inducible prophage. As expected, rhlR mutants of ID4365 behave as social cheaters, and exoprotease-deficient individuals accumulate upon continuous growth in casein. Moreover, in all continuous cultures, population collapses occur. However, this also sometimes happens before cheaters dominate. Interestingly, during growth in casein, ID4565's native prophage is induced, suggesting that the metabolic costs imposed by social cheating may increase its induction, promoting population collapses. Accordingly, lysogenization of the PAO1 lasR mutant with this prophage accelerated its collapse. These findings highlight the influence of temperate phages in social cheating.

Risk factors and outcomes of lower respiratory tract infections after traumatic brain injury: a retrospective observational study.

Background: Traumatic brain injury (TBI) is a public health problem with a high burden in terms of disability and death. Infections are a common complication, with respiratory infections being the most frequent. Most available studies have addressed the impact of ventilator-associated pneumonia (VAP) after TBI; therefore, we aim to characterize the hospital impact of a broader entity, lower respiratory tract infections (LRTIs). Methods: This observational, retrospective, single-center cohort study describes the clinical features and risk factors associated with LRTIs in patients with TBI admitted to an intensive care unit (ICU). We used bivariate and multivariate logistic regressions to identify the risk factors associated with developing LRTI and determine its impact on hospital mortality. Results: We included 291 patients, of whom 77% (225/291) were men. The median (IQR) age was 38 years (28-52 years). The most common cause of injury was road traffic accidents 72% (210/291), followed by falls 18% (52/291) and assault at 3% (9/291). The median (IQR) Glasgow Coma Scale (GCS) score on admission was 9 (6-14), and 47% (136/291) were classified as severe TBI, 13% (37/291) as moderate TBI, and 40% (114/291) as mild TBI. The median (IQR) injury severity score (ISS) was 24 (16-30). Nearly 48% (141/291) of patients presented at least one infection during hospitalization, and from those, 77% (109/141) were classified as LRTIs, which included tracheitis 55% (61/109), ventilator-associated pneumonia (VAP) 34% (37/109), and hospital-acquired pneumoniae (HAP) 19% (21/109). After multivariable analysis, the following variables were significantly associated with LRTIs: age (OR 1.1, 95% CI 1.01-1.2), severe TBI (OR 2.7, 95% CI 1.1-6.9), AIS thorax (OR 1.4, 95 CI 1.1-1.8), and mechanical ventilation on admission (OR 3.7, 95% CI 1.1-13.5). At the same time, hospital mortality did not differ between groups (LRTI 18.6% vs. No LRTI 20.1%, p = 0.7), and ICU and hospital length of stay (LOS) were longer in the LRTI group (median [IQR] 12 [9-17] vs. 5 [3-9], p < 0.01) and (median [IQR] 21 [13-33] vs. 10 [5-18], p = 0.01), respectively. Time on the ventilator was longer for those with LRTIs. Conclusion: The most common site/location of infection in patients with TBI admitted to ICU is respiratory. Age, severe TBI, thoracic trauma, and mechanical ventilation were identified as potential risk factors. LRTI was associated with prolonged ICU, hospital stay, and more days on a ventilator, but not with mortality.

Marginal versus Segmental Mandibulectomy in the Treatment of Oral Cavity Cancer: A Systematic Review and Meta-analysis

Abstract Introduction Oral cavity squamous cell carcinoma (OCSCC) is the most common malignancy in the oral cavity. Two types of mandibular resections have been described: the segmental mandibulectomy and the marginal mandibulectomy. Both may have a different impact over the quality of life, oncological prognosis, and functional or aesthetic result. Objectives The aim of this study was to systematically explore the literature to determine the survival outcomes and disease control rates in patients who underwent segmental or marginal mandibulectomy for OCSCC with histological evidence of cortical and medullary bone invasion. Data Synthesis This review involved a systematic search of the electronic databases MEDLINE/PUBMED, Google Scholar, Ovid Medline, Embase, and Scopus including articles from 1985 to 2019. Fifteen articles were included for qualitative analysis and 11 articles were considered for meta-analysis calculations. All of them correspond to retrospective cohort studies. Conclusion This systematic review reveals the low-level evidence regarding the impact over local control or survival according to the type of mandibulectomy. Our results need to be considered with precaution according to the limited evidence available. We just found difference regarding the 5-year disease-free survival, and a tendency in favor of segmental mandibulectomy was confirmed when medullary invasion was evident.

Pseudomonas aeruginosa LasR overexpression leads to a RsaL-independent pyocyanin production inhibition in a low phosphate condition.

Several Pseudomonas aeruginosa virulence-related traits like pyocyanin are regulated by an intricate regulatory network called quorum sensing (QS) that relies on transcriptional regulators that are activated through binding to a self-produced molecule called an autoinducer (AI). QS is composed of three systems, Las, Rhl and Pqs. In the Las system, the regulatory protein LasR interacts with its AI to activate the other two QS systems. In turn, the Rhl and Pqs systems regulate the expression of multiple virulence-related genes, such as the genes of the reiterated operons phzA1B1C1D1E1F1G1 and phzA2B2C2D2E2F2G2 involved in pyocyanin production. The Las system also regulates the negative regulator RsaL, which provides negative feedback to the QS-response, including repression of pyocyanin synthesis genes. In this work, we describe that LasR can act as a negative regulator of phzA1 transcription and hence of pyocyanin production and that this regulation is independent of RsaL activity. This work contributes to the understanding of QS-dependent pyocyanin production and demonstrates a previously uncharacterized role of LasR as a repressor.

COVID-19 Pandemic and its Impact on the Management of Head and Neck Cancer in the Spanish Healthcare System.

Introduction Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has represented a major challenge for healthcare systems worldwide, changing the habits of physicians. A reorganization of healthcare activity has been necessary, limiting surgical activity to essential cases (emergencies and oncology), and improving the distribution of health resources. Objective To analyze the impact of the COVID-19 pandemic on head and neck cancer surgery management in Spain. Methods A cross-sectional study, through an anonymous and voluntary online survey distributed to 76 Spanish otorhinolaryngology departments. Results A total of 44 centers completed the survey, 65.9% of which were high-volume. A total of 45.5% of them had to stop high-priority surgery and 54.5% of head and neck surgeons were relocated outside their scope of practice. Surgeons reported not feeling safe during their usual practice, with a decrease to a 25% of airway procedures. A total of 29.5% were "forced" to deviate from the "standard of care" due to the epidemiological situation. Conclusions Approximately half of the departments decreased their activity, not treating their patients on a regular basis, and surgeons were reassigned to other tasks. It seems necessary that the head and neck surgeons balance infection risk with patient care. The consequences of the reported delays and changes in daily practice should be evaluated in the future in order to understand the real impact of the pandemic on the survival of head and neck cancer patients.

PqsR-independent quorum-sensing response of Pseudomonas aeruginosa ATCC 9027 outlier-strain reveals new insights on the PqsE effect on RhlR activity.

Pseudomonas aeruginosa is a ubiquitous environmental bacterium and an opportunistic pathogen that represents an important health hazard. The quorum-sensing response regulates the expression of several virulence factors and involves three regulons: Las, Rhl, and Pqs. The P. aeruginosa ATCC 9027 strain, which belongs to the genetically diverse PA7 clade, contains a frame-shift mutation in the pqsR gene that encodes a transcriptional activator necessary for pyocyanin (PYO) synthesis in type strains PAO1 and PA14. Here we characterize the PqsE-dependent production of PYO in strain ATCC 9027. We show that this strain expresses pqsE independently of PqsR and in the absence of quinolone production, and that PqsE promotes the RhlR-dependent production of PYO, yet this production is not strictly dependent on PqsE. In addition, we show that in both strains ATCC 9027 and PAO1, PqsE overexpression causes an increased concentration of RhlR and enhances PYO production but does not affect rhamnolipids (RL) production in the same way. These results suggest that PqsE interaction with RhlR preferentially modifies its ability to activate transcription of genes involved in PYO production and provide new evidence about PqsE-dependent RhlR activation, highlighting the variability of the QS response among different P. aeruginosa clades and strains. HIGHLIGHTS: Pseudomonas aeruginosa ATCC 9027 is able to produce pyocyanin in phosphate limiting conditions, even in the absence of a functional PqsR. This strain does not produce alkyl quinolones like PQS and HHQ, but expresses pqsE. Synthesis of pyocyanin by ATCC 9027 is only partially dependent on pqsE. The overexpression of pqsE in the ATCC 9027 and PAO1 strains causes pyocyanin overproduction. The overexpression of pqsE in these strains causes an increased RhlR concentration without affecting rhlR transcription or translation. Rhamnolipids production is not affected to the same extent as pyocyanin by overexpression of pqsE in these strains.

Assessment of mandibular kinematics values and its relevance for the diagnosis of temporomandibular joint disorders.

BACKGROUND/PURPOSE: Temporomandibular disorders (TMD) are common conditions that involve the temporomandibular joints (TMJs), jaw muscles, or both, and can cause alteration in the mandibular kinematics. The aim of this study was to assess the relationship between mandibular kinematics and temporomandibular joint disorders (TMJD) as a clinical tool for evaluation and diagnosis of these patients. MATERIALS AND METHODS: A retrospective study based on the analysis of the clinical findings from patients' charts was carried out, with a sample size of 476 patients. Statistical analysis was made with chi-square test for qualitative variables and student t-test for quantitative variables. Then, odds ratio with its confidence interval were calculated. A p value < 0.05 was considered statistically significant. RESULTS: Most patients were female (80.7%) and between 16 and 25 years old. Disc displacement with reduction (DDwR) and subluxation were associated with increased kinematic parameters, while disc displacement without reduction (DDwoR) and retrodiscitis were associated with decreased kinematic values. A soft end feel was related to osteoarthritis (OA). Structural incompatibility was most prevalent in older patients. CONCLUSION: Mandibular kinematic values are associated with specific temporomandibular joint disorders and could be considered as a useful clinical tool to perform the right diagnosis of TMJD.

Cyclic di-GMP-Mediated Regulation of Extracellular Mannuronan C-5 Epimerases Is Essential for Cyst Formation in Azotobacter vinelandii.

The genus Azotobacter, belonging to the Pseudomonadaceae family, is characterized by the formation of cysts, which are metabolically dormant cells produced under adverse conditions and able to resist desiccation. Although this developmental process has served as a model for the study of cell differentiation in Gram-negative bacteria, the molecular basis of its regulation is still poorly understood. Here, we report that the ubiquitous second messenger cyclic dimeric GMP (c-di-GMP) is critical for the formation of cysts in Azotobacter vinelandii Upon encystment induction, the levels of c-di-GMP increased, reaching a peak within the first 6 h. In the absence of the diguanylate cyclase MucR, however, the levels of this second messenger remained low throughout the developmental process. A. vinelandii cysts are surrounded by two alginate layers with variable proportions of guluronic residues, which are introduced into the final alginate chain by extracellular mannuronic C-5 epimerases of the AlgE1 to AlgE7 family. Unlike in Pseudomonas aeruginosa, MucR was not required for alginate polymerization in A. vinelandii Conversely, MucR was necessary for the expression of extracellular alginate C-5 epimerases; therefore, the MucR-deficient strain produced cyst-like structures devoid of the alginate capsule and unable to resist desiccation. Expression of mucR was partially dependent on the response regulator AlgR, which binds to two sites in the mucR promoter, enhancing mucR transcription. Together, these results indicate that the developmental process of A. vinelandii is controlled through a signaling module that involves activation by the response regulator AlgR and c-di-GMP accumulation that depends on MucR.IMPORTANCEA. vinelandii has served as an experimental model for the study of the differentiation processes to form metabolically dormant cells in Gram-negative bacteria. This work identifies c-di-GMP as a critical regulator for the production of alginates with specific contents of guluronic residues that are able to structure the rigid laminated layers of the cyst envelope. Although allosteric activation of the alginate polymerase complex Alg8-Alg44 by c-di-GMP has long been recognized, our results show a previously unidentified role during the polymer modification step, controlling the expression of extracellular alginate epimerases. Our results also highlight the importance of c-di-GMP in the control of the physical properties of alginate, which ultimately determine the desiccation resistance of the differentiated cell.

The third quorum-sensing system of Pseudomonas aeruginosa: Pseudomonas quinolone signal and the enigmatic PqsE protein.

Pseudomonas aeruginosa is an opportunistic pathogen that produces several virulence factors such as lectin A, pyocyanin, elastase and rhamnolipids. These compounds are controlled transcriptionally by three quorum-sensing circuits, two based on the synthesis and detection of N-acyl-homoserine-lactone termed the Las and Rhl system and a third system named the Pseudomonas quinolone signal (PQS) system, which is responsible for generating 2-alkyl-4(1 h)-quinolones (AQs). The transcriptional regulator called PqsR binds to the promoter of pqsABCDE in the presence of PQS or HHQ creating a positive feedback-loop. PqsE, encoded in the operon for AQ synthesis, is a crucial protein for pyocyanin production, activating the Rhl system by a still not fully understood mechanism. In turn, the regulation of the PQS system is modulated by Las and Rhl systems, which act positively and negatively, respectively. This review focuses on the PQS system, from its discovery to its role in Pseudomonas pathogenesis, such as iron depletion and pyocyanin synthesis that involves the PqsE protein - an intriguing player of this system.

Assessment of association between muscular diagnosis in temporomandibular disorders with mandibular kinematics.

Objective: To study the association between mandibular kinematics and muscular diagnoses in temporomandibular disorders (TMD). Method: Retrospective, descriptive clinical study based on an analysis of 370 clinical records of muscle diagnoses in TMD. Statistical analysis was performed with the SPSS program. Results: Five different muscle diagnoses were considered. Myospasm and centrally mediated myalgia were discarded from the statistical analysis because of their low incidences. The diagnoses included in the analysis were protective co-contraction, myofascial pain, and myalgia. Decreased mandibular kinematics in the frontal and horizontal planes were related to myofascial pain. Mandibular kinematics with deflection in opening values decreased in the frontal plane, but intermediate or increased values in the horizontal plane were related to the protective co-contraction. Muscle pain with normal or increased mandibular kinematics was associated with a diagnosis of myalgia. Conclusion: Values of mandibular kinematics are correlated with the muscular diagnoses in TMD and are useful for diagnosis.

Morphological analysis of Trichomycterus areolatus Valenciennes, 1846 from southern Chilean rivers using a truss-based system (Siluriformes, Trichomycteridae).

Trichomycterus areolatus Valenciennes, 1846 is a small endemic catfish inhabiting the Andean river basins of Chile. In this study, the morphological variability of three T. areolatus populations, collected in two river basins from southern Chile, was assessed with multivariate analyses, including principal component analysis (PCA) and discriminant function analysis (DFA). It is hypothesized that populations must segregate morphologically from each other based on the river basin that they were sampled from, since each basin presents relatively particular hydrological characteristics. Significant morphological differences among the three populations were found with PCA (ANOSIM test, r = 0.552, p < 0.0001) and DFA (Wilks's λ = 0.036, p < 0.01). PCA accounted for a total variation of 56.16% by the first two principal components. The first Principal Component (PC1) and PC2 explained 34.72 and 21.44% of the total variation, respectively. The scatter-plot of the first two discriminant functions (DF1 on DF2) also validated the existence of three different populations. In group classification using DFA, 93.3% of the specimens were correctly-classified into their original populations. Of the total of 22 transformed truss measurements, 17 exhibited highly significant (p < 0.01) differences among populations. The data support the existence of T. areolatus morphological variation across different rivers in southern Chile, likely reflecting the geographic isolation underlying population structure of the species.

Pseudomonas aeruginosa ATCC 9027 is a non-virulent strain suitable for mono-rhamnolipids production.

Rhamnolipids produced by Pseudomonas aeruginosa are biosurfactants with a high biotechnological potential, but their extensive commercialization is limited by the potential virulence of P. aeruginosa and by restrictions in producing these surfactants in heterologous hosts. In this work, we report the characterization of P. aeruginosa strain ATCC 9027 in terms of its genome-sequence, virulence, antibiotic resistance, and its ability to produce mono-rhamnolipids when carrying plasmids with different cloned genes from the type strain PAO1. The genes that were expressed from the plasmids are those coding for enzymes involved in the synthesis of this biosurfactant (rhlA and rhlB), as well as the gene that codes for the RhlR transcriptional regulator. We confirm that strain ATCC 9027 forms part of the PA7 clade, but contrary to strain PA7, it is sensitive to antibiotics and is completely avirulent in a mouse model. We also report that strain ATCC 9027 mono-rhamnolipid synthesis is limited by the expression of the rhlAB-R operon. Thus, this strain carrying the rhlAB-R operon produces similar rhamnolipids levels as PAO1 strain. We determined that strain ATCC 9027 with rhlAB-R operon was not virulent to mice. These results show that strain ATCC 9027, expressing PAO1 rhlAB-R operon, has a high biotechnological potential for industrial mono-rhamnolipid production.

Use of tree species by White-throated treerunner (Pygarrhichas albogularis King) in a secondary native forest of southern Chile / Uso de especies de árboles por el Comesebo Grande (Pygarrhichas albogularis King) en un bosque nativo secundario del sur de Chile

En ecosistemas boscosos, numerosas especies de aves insectívoras usan ciertas especies de árboles como sustrato de alimentación y de nidificación. Evaluamos el uso de Pygarrhichas albogularis King, 1831 por componentes florísticos utilizados como sustrato de alimentación en un bosque nativo en el sur de Chile entre el 2009 y 2010. De 13 especies de árboles y arbustos P. albogularis utilizó seis especies de árboles como sustrato de alimentación. El uso de los árboles se limitó a alturas intermedias (11-20 m) y principalmente utilizó el tronco (40% observaciones) y las ramas secundarias (26%) de los árboles como sustrato de alimentación. Pygarrhichas albogularis mostró un uso desproporcionado por Nothofagus dombeyi y un uso importante por árboles con la mayor estructura etárea (DAP 81-100 cm). Por su parte, N. dombeyi presentó una profundidad de las grietas de la corteza significativamente mayor que E. cordifolia. A su vez, la profundidad de las grietas covarió positiva y significativamente con la oferta de invertebrados en la corteza de los árboles. Consideramos la profundidad de la corteza y la oferta de invertebrados como las causas próximas que podrían explicar el uso desproporcionado de P. albogularis por Nothofagus dombeyi.(AU)

In forest ecosystems, numerous species of insectivorous birds use certain tree species as feeding and nesting substrates. Between 2009 and 2010, the use of different floristic components as feeding substrate by the Pygarrhichas albogularis King, 1831 was evaluated in a southern Chilean secondary native forest. From a total of 13 trees and bush species, six tree species were used by P. albogularis as a feeding substrate. Tree use was limited to intermediate heights (11-20 m) and, mainly, to the trunk (40% of observations) and secondary branches (26%). Pygarrhichas albogularis showed a disproportionated use of N. dombeyi and an important use of trees with a greater age structure (DBH 81-100 cm). Nothofagus dombeyi presented a significantly greater tree bark crevice depth than E. cordifolia. In turn, covariance between crevice depth and invertebrate supply in tree bark was positive and significant. We consider bark depth and invertebrate supply to be the proximate causes explaining P. albogularis disproportionated use of Nothofagus dombeyi.(AU)

Use of tree species by White-throated treerunner (Pygarrhichas albogularis King) in a secondary native forest of southern Chile / Uso de especies de árboles por el Comesebo Grande (Pygarrhichas albogularis King) en un bosque nativo secundario del sur de Chile

En ecosistemas boscosos, numerosas especies de aves insectívoras usan ciertas especies de árboles como sustrato de alimentación y de nidificación. Evaluamos el uso de Pygarrhichas albogularis King, 1831 por componentes florísticos utilizados como sustrato de alimentación en un bosque nativo en el sur de Chile entre el 2009 y 2010. De 13 especies de árboles y arbustos P. albogularis utilizó seis especies de árboles como sustrato de alimentación. El uso de los árboles se limitó a alturas intermedias (11-20 m) y principalmente utilizó el tronco (40% observaciones) y las ramas secundarias (26%) de los árboles como sustrato de alimentación. Pygarrhichas albogularis mostró un uso desproporcionado por Nothofagus dombeyi y un uso importante por árboles con la mayor estructura etárea (DAP 81-100 cm). Por su parte, N. dombeyi presentó una profundidad de las grietas de la corteza significativamente mayor que E. cordifolia. A su vez, la profundidad de las grietas covarió positiva y significativamente con la oferta de invertebrados en la corteza de los árboles. Consideramos la profundidad de la corteza y la oferta de invertebrados como las causas próximas que podrían explicar el uso desproporcionado de P. albogularis por Nothofagus dombeyi.

In forest ecosystems, numerous species of insectivorous birds use certain tree species as feeding and nesting substrates. Between 2009 and 2010, the use of different floristic components as feeding substrate by the Pygarrhichas albogularis King, 1831 was evaluated in a southern Chilean secondary native forest. From a total of 13 trees and bush species, six tree species were used by P. albogularis as a feeding substrate. Tree use was limited to intermediate heights (11-20 m) and, mainly, to the trunk (40% of observations) and secondary branches (26%). Pygarrhichas albogularis showed a disproportionated use of N. dombeyi and an important use of trees with a greater age structure (DBH 81-100 cm). Nothofagus dombeyi presented a significantly greater tree bark crevice depth than E. cordifolia. In turn, covariance between crevice depth and invertebrate supply in tree bark was positive and significant. We consider bark depth and invertebrate supply to be the proximate causes explaining P. albogularis disproportionated use of Nothofagus dombeyi.

A small heat-shock protein (Hsp20) regulated by RpoS is essential for cyst desiccation resistance in Azotobacter vinelandii.

In Azotobacter vinelandii, a cyst-forming bacterium, the alternative sigma factor RpoS is essential to the formation of cysts resistant to desiccation and to synthesis of the cyst-specific lipids, alkylresorcinols. In this study, we carried out a proteome analysis of vegetative cells and cysts of A. vinelandii strain AEIV and its rpoS mutant derivative AErpoS. This analysis allowed us to identify a small heat-shock protein, Hsp20, as one of the most abundant proteins of cysts regulated by RpoS. Inactivation of hsp20 did not affect the synthesis of alkylresorcinols or the formation of cysts with WT morphology; however, the cysts formed by the hsp20 mutant strain were unable to resist desiccation. We also demonstrated that expression of hsp20 from an RpoS-independent promoter in the AErpoS mutant strain is not enough to restore the phenotype of resistance to desiccation. These results indicate that Hsp20 is essential for the resistance to desiccation of A. vinelandii cysts, probably by preventing the aggregation of proteins caused by the lack of water. To our knowledge, this is the first report of a small heat-shock protein that is essential for desiccation resistance in bacteria.

Roles of RpoS and PsrA in cyst formation and alkylresorcinol synthesis in Azotobacter vinelandii.

Azotobacter vinelandii is a soil bacterium that undergoes differentiation to form cysts that are resistant to desiccation. Upon induction of cyst formation, the bacterium synthesizes alkylresorcinols that are present in cysts but not in vegetative cells. Alternative sigma factors play important roles in differentiation. In A. vinelandii, AlgU (sigma E) is involved in controlling the loss of flagella upon induction of encystment. We investigated the involvement of the sigma factor RpoS in cyst formation in A. vinelandii. We analysed the transcriptional regulation of the rpoS gene by PsrA, the main regulator of rpoS in Pseudomonas species, which are closely related to A. vinelandii. Inactivation of rpoS resulted in the inability to form cysts resistant to desiccation and to produce cyst-specific alkylresorcinols, whereas inactivation of psrA reduced by 50 % both production of alkylresorcinols and formation of cysts resistant to desiccation. Electrophoretic mobility shift assays revealed specific binding of PsrA to the rpoS promoter region and that inactivation of psrA reduced rpoS transcription by 60 %. These results indicate that RpoS and PsrA are involved in regulation of encystment and alkylresorcinol synthesis in A. vinelandii.