Brazilian Journal of Microbiology最新文献

The fact that SARS-CoV-2 has reportedly infected companion, livestock and wildlife animals may constitute a significant risk for virus reservoirs, ground for emerging variants and potential for novel reverse zoonosis. Hence, SARS-CoV-2 surveillance in animal species is crucial to prevent emerging variants which may spread to humans. The present study aimed to develop a simple, high-throughput and ultrafast magnetic bead immunoassay to detect anti-SARS-CoV-2 nucleocapsid and spike reactive IgG antibodies in dog and cat serum samples. The assays were validated using serum from eleven dogs and cats which had SARS-CoV-2 infections confirmed by real-time RT-PCR. The negative cohort consisted of pre-pandemic dog and cat samples. The assays performed at 73-82% sensitivity and 97.5-98% specificity for dogs and 71% sensitivity and 92-94% specificity for cats. The lower assay specificity for cats is explained by the fact that cat pre-pandemic sera showed high levels of cross-reactive with SARS-CoV-2 Nucleocapsid and Spike, supporting that these animals have been exposed to other coronavirus sharing structural similarities with SARS-CoV-2. These assays described in this work are now being used for SARS-CoV-2 surveillance and research purposes.

Antibiotics are misused nowadays, leading to the prevalence of antibiotic resistant bacterial strains; causing the world to move towards natural medicine. Retama raetam had wide medicinal use. In the present study, R. raetam ethanolic extract proved to be active against Pseudomonas aeruginosa with MIC values ranged from 15.62 to 250 µg/ml. Antioxidant analysis showed that the extract had high scavenging activity reached 92.40%. GC/MS analysis revealed that Sparteine and Tributyl acetylcitrate represent the extract major components. Furthermore, the combination between Retama raetam extract and colistin showed a synergistic effect. Moreover, nano-phytosome was designated and optimized to encapsulate Retama raetam extract/Colistin. Nano-phytosome characterized by particle size, Zeta potential, polydispersity index and Entrapment efficiency percentage of 16.92-32.85 nm, -30.40 mV, 0.26 and 89% respectively. The antibacterial activity of the prepared nano-phytosome formula against P. aeruginosa showed promising MIC, MBC, MIC index, and IZ diameter reaching 7.81, 15.62 µg/ml, 2, and 39 mm, respectively. While TEM examination of P. aeruginosa cells treated with nano-phytosome formula revealed cell wall breakage which led to cell death. Finally, P. aeruginosa LPS was used to induce neurodegenerative disease in rat model. Rats treated with nano-phytosome formula showed normal histoarchitecture organization and the cerebral cortex was partially restored compared to control groups.

Objectives

This study addressed the need for new treatments for severe Candida infections, especially resistant strains. It evaluated the antifungal potential of geraniol alone and with fluconazole against various Candida spp., including resistant strains, and investigated geraniol’s mechanism of action using flow cytometry.

Methods

The research assessed the inhibitory effects of geraniol on the growth of various Candida species at concentrations ranging from 110 to 883 µg/ml. The study also explored the potential synergistic effects when geraniol was combined with fluconazole. The mechanism of action was investigated through flow cytometry, with a particular emphasis on key enzymes associated with plasma membrane synthesis, membrane permeability changes, mitochondrial membrane depolarization, reactive oxygen species (ROS) induction, and genotoxicity.

Results

Geraniol demonstrated significant antifungal activity against different Candida species, inhibiting growth at concentrations within the range of 110 to 883 µg/ml. The mechanism of action appeared to be multifactorial. Geraniol was associated with the inhibition of crucial enzymes involved in plasma membrane synthesis, increased membrane permeability, induction of mitochondrial membrane depolarization, elevated ROS levels, and the presence of genotoxicity. These effects collectively contributed to cell apoptosis.

Conclusions

Geraniol, alone and in combination with fluconazole, shows promise as a potential therapeutic option for Candida spp. infections. Its diverse mechanism of action, impacting crucial cellular processes, highlights its potential as an effective antifungal agent. Further research into geraniol’s therapeutic applications may aid in developing innovative strategies to address Candida infections, especially those resistant to current therapies.

Many cooked foods are prepared with spices and dried herbs; these can be contaminated by several types of microorganisms, including aerobic spore-forming bacteria. The Bacillus cereus group is very widespread in nature and is known among the common food contaminants. They are involved in food poisoning, causing two types of syndromes, diarrheal and emetic. The aims of the present work were to determine the prevalence of toxigenic Bacillus cereus spores in spices and herbs marketed in the Laghouat area and to identify their toxigenic genes via PCR. Among the 191 samples, 14.13% were determined to be B. cereus, with concentrations ranging from 2.52 to 5.82 log cfu/g, where the highest level of contamination was observed in allspice and ginger. Moreover, entFM (100%), nhe (88.23%) and cytK (70.58%) were the most frequently identified toxin genes, whereas hbl (23.52%) was less common, and no emetic toxin-encoding gene (cesB) was found in any of the samples. Considering the results of the present study, the B. cereus microbial load and toxin gene profiles of spices show that spices have potential for public health in Algeria. In this context, it is crucial to guarantee the microbiological safety of spices by respecting good hygiene practices, eliminating bacterial spores and toxin production via sterilization and using appropriate packaging for these products.

Cold-adapted microorganisms can produce enzymes with activity at low and mild temperatures, which can be applied to environmental biotechnology. This study aimed to characterize 20 Antarctic fungi to identify their genus (ITS rDNA marker) and growth temperatures and evaluate their ability to decolorize and detoxify the textile dye indigo carmine (IC). An individual screening was performed to assess the decolorization and detoxification of IC by the isolates, as well as in consortia with other fungi. The isolates were affiliated with seven ascomycete genera: Aspergillus (n = 4), Cosmospora (n = 2), Leuconeurospora (n = 2), Penicillium (n = 3), Pseudogymnoascus (n = 6), Thelebolus (n = 2), and Trichoderma (n = 1). The two isolates from the genus Leuconeurospora were characterized as psychrophilic, while the others were psychrotolerant. The Penicillium isolates were able to decolorize between 60 and 82% of IC. The isolates identified as Pseudogymnoascus showed the best detoxification capacity, with results varying from 49 to 74%. The consortium using only Antarctic ascomycetes (C1) showed 45% of decolorization, while the consortia with the addition of basidiomycetes (C1 + Peniophora and C1 + Pholiota) showed 40% and 50%, respectively. The consortia C1 with the addition of the basidiomycetes presented a lower toxicity after the treatments. In addition, a higher fungal biomass was produced in the presence of dye when compared with the experiment without the dye, which can be indicative of dye metabolization. The results highlight the potential of marine-derived Antarctic fungi in the process of textile dye degradation. The findings encourage further studies to elucidate the degradation and detoxification pathways of the dye IC by these fungal isolates.

Finding novel promoter sequences is a cornerstone of synthetic biology. To contribute to the expanding catalog of biological parts, we employed a promoter-trap approach to identify novel sequences within an Antarctic microbial community that act as broad host-range promoters functional in diverse Pseudomonadota. Using Pseudomonas putida KT2440 as host, we generated a library comprising approximately 2,000 clones resulting in the identification of thirteen functional promoter sequences, thereby expanding the genetic toolkit available for this chassis. Some of the discovered promoter sequences prove to be broad host-range as they drove gene expression not only in P. putida KT2440 but also in Escherichia coli DH5α, Cupriavidus taiwanensis R1^T, Paraburkholderia phymatum STM 815^T, Ensifer meliloti 1021, and an indigenous Antarctic bacterium, Pseudomonas sp. UYIF39. Our findings enrich the existing catalog of biological parts, offering a repertoire of broad host-range promoter sequences that exhibit functionality across diverse members of the phylum Pseudomonadota, proving Antarctic microbial community as a valuable resource for prospecting new biological parts for synthetic biology.

Neutralizing antibody (nAb) responses against SARS-CoV-2 variants after inactivated virus vaccine (CoronaVac) in kidney transplant recipients (KTRs) with or without SARS-CoV-2 infection history remains unclear. We aimed to evaluate the neutralizing antibody responses against emerging SARS-CoV-2 variants after two doses of CoronaVac in these patients. 22.2% of participants had hybrid immunity. Anti-spike IgG antibodies were evidenced in 44% of the patients. nAbs against B.1.111, Mu, and Omicron were detected in 28.5%, 17.9%, and 21.4% of naïve KTRs, respectively. Furthermore, nearly 100% of KTRs with hybrid immunity had nAbs against the variants evaluated. Thus, a significant proportion of infection-naïve KTRs had no detectable nAb titers against Mu and Omicron variants after two doses of the CoronaVac vaccine. However, the nAb titers were significantly higher in patients with hybrid immunity, and it was no association between the immunosuppressive regimen and the seropositivity rate of anti-SARS-CoV-2 neutralizing antibodies. Therefore, hybrid KTRs are protected against COVID-19 by emerging variants able to escape from vaccine-elicited nAbs such as Mu and Omicron.

The use of pesticides, such as glyphosate, has increased due to population growth and the rising demand for food. Plant growth-promoting rhizobacteria (PGPR), such as Streptomyces, offer a more ecologically friendly alternative to the excessive use of pesticides. However, these bacteria undergo a complex life cycle involving the formation of hyphae, mycelia, and spores, which makes standardizing laboratory cultures challenging. In this context, we tested three methods for cultivating a Streptomyces isolate (CLV322) in the presence of the stressor agent glyphosate, denoted as M1, M2, and M3. These methods involved the simultaneous addition of the herbicide 24–48 h after the start of cultivation. We evaluated the growth and cell viability of CLV322 using the 2,3,5-triphenyl tetrazolium chloride (TTC) assay under glyphosate-based herbicide stress (Roundup^® Original DI) at concentrations ranging from 0.002 to 7.2 mg mL^− 1. We also assessed the ability of CLV322 to maintain PGPR characteristics in the presence of the herbicide by quantifying indolic compounds, siderophores, and phenazines. The cultivation method significantly influenced the production of metabolites by CLV322, with M3 yielding more consistent results across the evaluated parameters. Our findings suggest that germinating Streptomyces spores for 48 h before introducing glyphosate (M3) enables the analysis of bacterial tolerance to herbicide stress. This methodology may also apply to evaluate other abiotic stresses on Streptomyces strains.

Streptococcus pneumoniae is an important human pathogen that can colonize the respiratory tract of healthy individuals. The respiratory tract mucosa is thus the first barrier for this pathogen. In this study, we have tested three models of the respiratory epithelium with immune cells: (i) monolayer of A549 human lung epithelial cells, (ii) A549 + macrophages differentiated from the human monocytic THP-1 cell line (dMφ) and (iii) A549 + dMφ + dendritic cells differentiated from THP-1 (dDC) using a two-chamber system. Pneumococcal strains Rx1 (non-encapsulated) and BHN418 (serotype 6B) were incubated with the cells and secretion of IL-6, IL-8, IL-1β, TNF-α and IL-10 was evaluated. Overall, the models using co-cultures of A549 + dMφ and A549 + dMφ + dDC elicited higher levels of pro-inflammatory cytokines and the non-encapsulated strain elicited an earlier cytokine response. BHN418 pspA (pneumococcal surface protein A) and pspC (pneumococcal surface protein C) knockouts elicited similar cytokine secretion in the co-culture models, whereas BHN18 ply (pneumolysin) knockout induced much lower levels. The results are in accordance with the activation of the inflammasome by Ply. Finally, we evaluated pneumococcal extracellular vesicles (pEVs) in the co-culture models and observed secretion of pro-inflammatory cytokines in the absence of cytotoxicity. Since pEVs are being studied as vaccine candidate against pneumococcal infections, the co-cultures of A549 + dMφ and A549 + dMφ + dDC are simple models that could be used to evaluate pEV vaccine batches.

Mushroom associated microbes could be utilized to improve crop productivity providing nutrients, plant growth promoting substances, production of hydrolytic enzymes and protecting plant from biotic and abiotic stress. An endophyte designated as KUFC101 was isolated from fruit body of Pleurotus ostreatus and identified as Porostereum umbrinoalutaceum based on nuclear-rRNA gene sequence analysis. Growth in different culture media, metal tolerance, biochemical characterization and effect on chilli plant growth promotion were studied. The isolate showed best growth in Malt extract medium and least growth in synthetic media. It could tolerate toxic metals (Mg, Ca, Fe, Cu, Mn, Zn and Cd each at 100 ppm concentration). It produced amylase, cellulase, chitinase, pectinase, catecholate type of siderophore and indole acetic acid, and inhibited growth of Alternaria solani and Penicillium citrinum. It could colonize in the rhizosphere of chilli plant and influence growth of chilli plant by improving biomass and metabolite content. Porostereum umbrinoalutaceum KUFC101 could be utilized in formulation of biofertiliser under sustainable agricultural system.