Brazilian Journal of Microbiology最新文献

Enterococci are members of the microbiota of humans and other animals. They can also be found in the environment, associated with food, healthcare infections, and hospital settings. Due to their wide distribution, they are inserted in the One Health context. The selective pressure caused by the extensive use of antimicrobial agents in humans, animals, and agriculture has increased the frequency of resistance to various drugs among enterococcal species. CRISPR-Cas system, an important prokaryotic defense mechanism against the entry of mobile genetic elements, may prevent the acquisition of genes involved in antimicrobial resistance and virulence. This system has been increasingly used as a gene editing tool, which can be used as a way to recognize and inactivate genes of interest. Here, we conduct a review on CRISPR systems found in enterococci, considering their occurrence, structure and organization, mechanisms of action and use as a genetic engineering technology. Type II-A CRISPR-Cas systems were shown to be the most frequent among enterococcal species, and the orphan CRISPR2 was the most commonly found system (54.1%) among enterococcal species, especially in Enterococcus faecalis. Distribution of CRISPR systems varied among species. CRISPR systems had 1 to 20 spacers, with size between 23 and 37 bp and direct repeat sequences from 25 to 37 bp. Several applications of the CRISPR-Cas biotechnology have been described in enterococci, mostly in vitro, using this editing tool to target resistance- and virulence-related genes.

Biofilm-associated disorders contribute to elevated morbidity and death rates among patients. We propose synthesizing niosomal structures containing the antibiotics tetracycline and ampicillin ((Tet/Amp)-Nio) and investigating their impact on standard strains of S. aureus, K. pneumoniae, and P. aeruginosa. The antibacterial and anti-biofilm effects of synthesized niosomes against standard pathogenic bacterial strains were studied, and also its cytotoxic activity was investigated against human foreskin fibroblast (HFF) cell line. The optimal formulation (F2) had an average particle size of 196.90 ± 4.57 nm, a PDI of 0.223 ± 0.013, a Zeta-potential of -19.25 ± 1.19 mV, a %EE of 70.92 ± 1.75% for Tet and 58.34 ± 1.85% for Amp, and a %Release rate of 49.34 ± 1.78% for Tet and 62.67 ± 1.19% for Amp. The release of Tet and Amp drugs over 48 h was 47% and 61%, respectively, from the (Tet/Amp)-Nio formulation. Also, our findings demonstrated that the Tet/Amp)-Nio have potent antibacterial, anti-biofilm, and lower cytotoxic activity compared to the Tet + Amp. In addition, (Tet/Amp)-Nio can upregulate the expression level of matrix metallopeptidase 2 (MMP2) and matrix metallopeptidase 9 (MMP9) genes, which shows their great activity in the wound healing process. The findings of the current investigation suggest that (Tet/Amp)-Nio enhances its antibacterial and antibiofilm effects against S. aureus, P. aeruginosa, and K. pneumoniae isolates. These formulations may serve as a novel approach for targeted drug delivery.

The rapid expansion of broiler chicken production in Brazil has presented significant sanitation challenges within the poultry industry. Among these challenges, Salmonella enterica subsp. enterica serotype Heidelberg stands as a contributor to global salmonellosis outbreaks. This study analyzed 13 draft genomes of Salmonella Heidelberg isolated from the pre-slaughter broiler chickens farms in Brazil. By conducting in silico analysis of these genomes, the study investigated genome similarity based on single nucleotide polymorphisms (SNPs) and identified genes encoding resistance to antimicrobials, sanitizers, and virulence factors. Furthermore, mobile genetic elements (MGE) were identified to assess their potential role in propagating genes through horizontal gene transfer. A risk classification was also applied based on the resistomes. The genomes revealed a high prevalence of genes conferring resistance to aminoglycosides, fosfomycin, sulfonamides, tetracycline, and genes linked to quaternary ammonium resistance. The study also uncovered six Salmonella pathogenicity islands (SPI) and over 100 genes encoding virulence factors. The association of MGE with antibiotic-resistant genes sul2 and blaCMY-2 raised concerns about the potential transfer to other bacteria, posing a substantial risk for spreading resistance mechanisms according to established risk protocols. Additionally, SNP analysis indicated close phylogenetic relationships among some isolates, suggesting a common origin. This study enhances our understanding of Salmonella Heidelberg strains by identifying key risk factors for transmission and revealing the association between resistance genes and MGEs. This insight provides a foundation for developing and implementing effective control, monitoring, and treatment strategies in the poultry industry.

Data on microbial fouling of various synthetic polymer materials, including those used in space technology, are summarized. It has been established that the dominant groups of microbiota of polymer fouling are the genera of mitosporous fungi Aspergillus, Penicillium, Alternaria, Trichoderma. The enzymatic properties of fungal strains from the collection of microbial cultures of the Microbial Depository Center of the National Academy of Sciences of Armenia were studied. It has been shown that Aspergillus fumigatus, Penicillium chrysogenum, P. steckii, Juxtiphoma eupyrena and a number of other fungi have biofouling activity towards polyethylene, polyethylene terephthalate and some other synthetic polymers. New fungal kits have been developed and proposed to evaluate the fungal resistance of polymeric materials. They include fungi isolated from bio-damaged polymers used in space technology and contain 2 to 5 fungal strains instead of 7 to 9 strains in previously used kits. Taking into account the obtained data, a comparative assessment of the fungal resistance of samples of synthetic polymeric materials of various classes that passed accelerated climatic tests has been carried out. It has been established that the kits of biodegradant fungi, composed of cultures of bio-damaged space technology, generally exceeded the activity of the previously used kits, based on which one can judge the obvious advantages of strains isolated from bio-damaged space technology. In the future, these kits could find application not only for biodegradation of polymers, but also for testing the biostability of various polymers, to use for the construction of aviation and space techniques. Moreover, new optimized kits may be developed based on the strains involved in this study.

This study aimed to assess the impact of S. aureus as well as Non-aureus Staphylococci and Mammaliicocci (NASM) subclinical mastitis-causing in the economic return in dairy herds. Data were gathered from the databases of five dairy herds located in the Agreste region of Pernambuco state, Northeastern Brazil, over a period of three consecutive months. A total of 155 mammary quarters from 155 healthy cows were categorized into the healthy group. Meanwhile, 257 mammary quarters from 187 cows that tested positive for either S. aureus or NASM were categorized into the infected group. The effect of mammary quarter infection in economic return was estimated using milk payment criteria on milk samples from healthy vs. infected cows based on a linear mixed model. Milk yield and milk price influenced the economic return, and both varied according to factors like herd, parity, stage of lactation, period of analysis, and the type of pathogen causing subclinical mastitis. There was a reduction in the average economic return caused by NASM infection (by 0.41 to 0.65 US$/cow/day) and S. aureus infection (by 0.25 to 0.36 US$/cow/day), when considering the effect of the pathogen over time for 1 and ≥ 2 infected quarters. Although no significant differences were observed in economic return between healthy cows and those infected with NASM or S. aureus, it is important to collect data on these pathogens for a more precise assessment of the economic impact of subclinical mastitis and for developing enhanced approaches for prevention and control.

Fusarium wilt is one of main phytopathology attacking tomato (Solanum lycopersicum L.) plantations in Brazil. Plant rhizosphere and endophytic beneficial microorganism are well known as plant growth promoters and biocontrol agents. The present study aims to evaluate the potential of different Bacillus strains as biocontrol agent to Fusarium oxysporum f. sp. lycopersici Race 3 strains; and also as plant growth promoting bacteria on Solanum lycopersicum cv Perinha. Different in vitro and greenhouse experiments were carried out to evaluate the direct and indirect bacterial-fungus antagonism, and they inoculation effects on plant traits. In vitro direct, metabolites, and volatile antagonism analysis demonstrated that B. toyonensis BR 10491(FORT 02) presented a broad antagonism to all tested race 3 FOL strains while B. megaterium BR 10466 (FORT 12), B. aryabhattai BR 10494 (FORT 25), B. stratosphericus BR 10438 (FORT 29) and B. cereus BR 10493 (FORT 113.1) strains showed significant antagonistic activity for at least two applied methods. Greenhouse pot experiments demonstrated a significant BCA effect of FORT 113.1 and FORT 02 against FOL Race 3 Fus 1302 strain during different tomato development stages (seedling, vegetative, and reproductive). Bacillus cereus (FORT 113.1) showed significantly higher shoot and height fresh weight, Chlorophyll a and Chlorophyll b content, stomata conductance, water use efficiency, and also a lower xylem infection percentage during vegetative and reproductive stages. Antioxidant enzymatic components analysis demonstrated a synergic effect of Fusarium and Bacillus inoculation, leading to a higher superoxide dismutase (SOD), catalase (CAT), and ascorbate peroxidase (APX) activity. In conclusion, the results suggest that strain FORT113.1 could be considered as a good candidate for production of new biofungicide with high potential to augment the existing biocontrol strategies.

Zika virus (ZIKV), a flavivirus, is one of the most serious re-emerging pathogens. Growing outbreaks in the Americas have linked ZIKV to significant clinical symptoms including Guillain-Barré syndrome in adults and congenital anomalies in newborns. ZIKV affects brain cells in a variety of ways, mostly apoptosis and cell cycle delays. Modulation of the host's immune reaction and the inflammatory process has also been shown to be involved in ZIKV-induced neurological disorders. This review summarized and discussed the latest advances in ZIKV research to shed fresh light on the multiple mechanisms incolved in ZIKV-induced neuropathogenesis.

Despite numerous studies on Salmonella enterica subsp. enterica serovar Typhimurium, the underlying mechanisms of several aspects of its virulence are still under investigation, including the role of the pdu and ttrA genes, associated with the metabolism of 1,2-propanediol using tetrathionate as an electron acceptor respectively. Our objective was to contribute to an understanding of the role of these genes inbacterial virulence for mice (Mus musculus) using an S. Typhumirum ΔttrApduA mutant. The experiment was conducted with a group infected by the S. Typhimurium mutant and a control group infected with a wild-type strain. The mutant was not attenuated compared with the parent strain. There were no differences in the bacterial numbers recovered from the mesenteric lymph nodes and Peyer's patches but at 8-day after oral infection higher numbers were recovered from the spleen, liver, and cecum. Unlike the single pduA and ttrA mutants, the double ΔttrApduA mutation did not affect invasion and survival in mice, which highlights the need for further studies to clarify the role of these important metabolism genes under reduced redox conditions linked to Salmonella virulence.

In the search for more sustainable remediation strategies for PAH-contaminated soils, an integrated application of thermal remediation and bioremediation (TEB) may allow the use of less impacting temperatures by associating heating to biological degradation. However, the influence of heating on soil microbiota remains poorly understood, especially in soils from tropical regions. This work investigated the effects of low-temperature heating on creosote-contaminated soil bacteria. We used culture-dependent and 16 S rRNA sequencing methods to compare the microbial community of soil samples heated to 60 and 100 ^oC for 1 h in microcosms. Heating to 60 °C reduced the density of cultivable heterotrophic bacteria compared to control soil (p < 0.05), and exposure to 100 °C inactivated the viable heterotrophic community. Burkholderia-Caballeronia-Paraburkholderia (BCP) group and Sphingobium were the predominant genera. Temperature and incubation time affected the Bray-Curtis dissimilarity index (p < 0.05). At 60 °C and 30 days incubation, the relative abundance of Sphingobium decreased and BCP increased dominance. The network of heated soil after 30 days of incubation showed fewer nodes and edges but maintained its density and complexity. Both main genera are associated with PAH degradation, suggesting functional redundancy and a likely potential of soil microbiota to maintain biodegradation ability after exposure to higher temperatures. We concluded that TEB can be considered as a potential strategy to bioremediate creosote-contaminated soils, allowing biodegradation in temperature ranges where thermal remediation does not completely remove contaminants. However, we recommend further research to determine degradation rates with this technology.

Silver nanoparticles (AgNPs) synthesized through green synthesis routes are widely used as antimicrobial agents due to their advantages such as biocompatibility, stability, sustainability, speed and cost-effectiveness. Although AgNPs appear to be more potent than silver ions, the mechanisms related to their antibacterial activity are not yet fully understood. The most common proposed mechanism of AgNPs' toxicity so far is the release of silver ions and/or specific functions of the particles. In this context, the present study aimed to investigate the mechanisms of action of AgNPs synthesized using noni fruit peels (Morinda citrifolia) against the phytopathogen Xanthomonas campestris pv. campestris (Xcc) through proteomics. Xcc was treated with AgNPs (32 µM), AgNO₃ (32 µM), or received no treatment (Ctrl - control condition), and its proteomic response was comprehensively characterized to elucidate the antimicrobial mechanisms of AgNPs in the phytopathogenic microorganism. A total of 352 differentially abundant proteins were identified. Most proteins were regulated in the AgNPs × Ctrl and AgNPs × AgNO₃ comparisons/conditions. When Xcc treated with 32 µM AgNPs were compared to controls, the results showed 134 differentially abundant proteins, including 107 increased and 27 decreased proteins. In contrast, when Xcc treated with 32 µM AgNO₃ were compared to Ctrl, the results showed only 14 differentially abundant proteins, including 10 increased proteins and 4 decreased proteins. Finally, when Xcc treated with 32 µM AgNPs were compared to Xcc treated with 32 µM AgNO₃, the results showed 204 differentially abundant proteins, including 75 increased proteins and 129 decreased proteins. Gene ontology enrichment analysis revealed that most of the increased proteins were involved in important biological processes such as metal ion homeostasis, detoxification, membrane organization, metabolic processes related to amino acids and carbohydrates, lipid metabolic processes, proteolysis, transmembrane transport, and others. The AgNPs used in this study demonstrated effective antimicrobial activity against the phytopathogenic bacteria Xcc. Furthermore, the obtained results contribute to a better understanding of the mechanisms of action of AgNPs in Xcc and may aid in the development of strategies to control Xcc in brassica.