ABSTRACT
The RNA chaperone Hfq promotes the association of small RNAs (sRNAs) with cognate mRNAs, controlling the expression of bacterial phenotype. Actinobacillus pleuropneumoniae hfq mutants strains are attenuated for virulence in pigs, impaired in the ability to form biofilms, and more susceptible to stress, but knowledge of the extent of sRNA involvement is limited. Here, using A. pleuropneumoniae strain MIDG2331 (serovar 8), 14 sRNAs were identified by co-immunoprecipitation with Hfq and the expression of eight, identified as trans-acting sRNAs, were confirmed by Northern blotting. We focused on one of these sRNAs, named Rna01, containing a putative promoter for RpoE (stress regulon) recognition. Knockout mutants of rna01 and a double knockout mutant of rna01 and hfq, both had decreased biofilm formation and hemolytic activity, attenuation for virulence in Galleria mellonella, altered stress susceptibility, and an altered outer membrane protein profile. Rna01 affected extracellular vesicle production, size and toxicity in G. mellonella. qRT-PCR analysis of rna01 and putative cognate mRNA targets indicated that Rna01 is associated with the extracytoplasmic stress response. This work increases our understanding of the multilayered and complex nature of the influence of Hfq-dependent sRNAs on the physiology and virulence of A. pleuropneumoniae.
ABSTRACT
The teaching-learning process becomes more attractive when practical classes are used as part of methodological tools. Aiming (i) to stimulate the interest of high school students in the microbiological world and (ii) to provide didactic experience for microbiology graduate students, practical classes were undertaken. These classes were carried out during the years 2016 and 2017, using the infrastructure held at Universidade Federal de Viçosa. Briefly, laboratory classes were prepared and taught by microbiology graduate students and an evaluation questionnaire was answered by the participants after the classes. Approximately 95% of the high school students, from both years, evaluated the experience outside of the school routine as very good and good. A total of 90.09% (2016) and 100% (2017) of graduate students evaluated this experience as very good and good. The relationship between high school and graduate students also increased the curiosity of the former regarding the university environment. Accordingly, the information retention regarding the 'microbiology world' was verified after one year and the students were able to remember important terms related to the microbiology class. In addition, this work allowed graduate students and high school students to build a closer relationship and created an excellent teaching-learning strategy for both.
ABSTRACT
The RNA chaperone Hfq regulates diverse processes in numerous bacteria. In this study, we compared phenotypes (growth rate, adherence, response to different stress conditions and virulence in Galleria mellonella) of wild-type (WT) and isogenic hfq mutants of three serovars (1, 8 and 15) of the porcine pathogen Actinobacillus pleuropneumoniae. Similar growth in rich broth was seen for all strains except Ap1∆hfq, which showed slightly reduced growth throughout the 24 h time course, and the complemented Ap8∆hfqC mutant had a prolonged lag phase. Differences were seen between the three serovar WT strains regarding adherence, stress response and virulence in G. mellonella, and deletion of hfq affected some, but not all of these phenotypes, depending on serovar. Complementation by expression of cloned hfq from an endogenous promoter only restored some WT phenotypes, indicating that complex regulatory networks may be involved, and that levels of Hfq may be as important as presence/absence of the protein regarding its contribution to gene regulation. Our results support that Hfq is a pleiotropic global regulator in A. pleuropneumoniae, but serovar-related differences exist. These results highlight the importance of testing multiple strains/serovars within a given species when determining contributions of global regulators, such as Hfq, to expression of complex phenotypes.
Subject(s)
Actinobacillus pleuropneumoniae/pathogenicity , Bacterial Adhesion , Host Factor 1 Protein/metabolism , Stress, Physiological , Virulence , Actinobacillus Infections/microbiology , Actinobacillus pleuropneumoniae/classification , Animals , Disease Models, Animal , Gene Deletion , Gene Expression Regulation, Bacterial , Genetic Complementation Test , Host Factor 1 Protein/genetics , Larva/microbiology , Moths/microbiology , Phenotype , Promoter Regions, Genetic , Serogroup , SwineABSTRACT
The presence of ants (Hymenoptera: Formicidae) in hospital environments may constitute a public health problem, especially since they are mechanical vectors for pathogenic organisms. This study aimed to survey the ant populations and analyze the presence of bacteria associated with them in two medium-sized regional hospitals in the municipality of Divinópolis, Minas Gerais, Brazil. Specimens were collected every monthly over a six-month period. The following ant species were found: Pheidole sp1 and sp2, Linepithema humile, Wasmannia auropunctata, Camponotus sp1 and sp2, Odontomachus sp, Solenopsis sp, Acromyrmex sp and Tapinoma melenocephalum. It was observed that these ants mechanically transported Pseudomonas aeruginosa, Enterococcus, Streptococcus, Escherichia coli and non-pathogenic and pathogenic Staphylococcus. These results show the propensity for occurrences of hospital infections at these sites caused by mechanical transmission of pathogens by ants.
Subject(s)
Ants/microbiology , Gram-Negative Bacteria/isolation & purification , Gram-Positive Bacteria/isolation & purification , Hospitals , Insect Vectors/microbiology , Animals , Ants/classification , Brazil , Insect Vectors/classificationABSTRACT
A presença de formigas (Hymenoptera: Formicidae) em ambientes hospitalares pode constituir um problema de saúde pública, especialmente por serem vetores mecânicos de organismos patogênicos. O trabalho teve como objetivo realizar o levantamento de formigas e analisar a presença de bactérias a elas associadas em dois hospitais regionais de médio porte da cidade de Divinópolis, MG. As coletas foram realizadas mensalmente, durante um período de seis meses. Foram coletadas formigas Pheidole sp1 e sp2, Linepithema humile, Wasmannia auropunctata, Camponotus sp1 e sp2, Odontomachus sp, Solenopsis sp, Acromyrmex sp e Tapinoma melenocephalum. Observou-se que estas transportavam mecanicamente Pseudomonas aeruginosa, Enterococcus, Streptococcus, Staphylococcus patogênico e não patogênico e Escherichia coli. Tais resultados evidenciam a propensão à ocorrência de infecções hospitalares nesses locais pela transmissão mecânica de agentes patogênicos por formigas.
The presence of ants (Hymenoptera: Formicidae) in hospital environments may constitute a public health problem, especially since they are mechanical vectors for pathogenic organisms. This study aimed to survey the ant populations and analyze the presence of bacteria associated with them in two medium-sized regional hospitals in the municipality of Divinópolis, Minas Gerais, Brazil. Specimens were collected every monthly over a six-month period. The following ant species were found: Pheidole sp1 and sp2, Linepithema humile, Wasmannia auropunctata, Camponotus sp1 and sp2, Odontomachus sp, Solenopsis sp, Acromyrmex sp and Tapinoma melenocephalum. It was observed that these ants mechanically transported Pseudomonas aeruginosa, Enterococcus, Streptococcus, Escherichia coli and non-pathogenic and pathogenic Staphylococcus. These results show the propensity for occurrences of hospital infections at these sites caused by mechanical transmission of pathogens by ants.
