RESUMO
A wide range of bioactive compounds with potential medical applications are produced by members of the genus Streptomyces. A new actinomycete producer of the antibiotic γ-rubromycin, designated TA 36, was isolated from an alpine soil sample collected in Peru (Machu Picchu). Morphological, physiological and biochemical characteristics of the strain, together with data obtained via phylogenetic analysis and MALDI-TOF MS, were used for the correct identification of the isolate. The isolate TA 36 showed morphological characteristics that were consistent with its classification within the genus Streptomyces. Phylogenetic analysis based on 16S rRNA gene sequences showed that the TA 36 strain was most similar to S. iakyrus and S. violaceochromogenes with 99% similarity. Phylogenetic analysis together with the profile of whole cell proteins indicated that the strain tested could be identified as S. iakyrus TA 36. The crude extract Ext.5333.TA 36 showed various effects against the tested organisms with strong antimicrobial activity in the growth of Staphylococcus aureus (Newman) (MIC value of 0.00195 µg/µL). HPLC fractionation and LC/MS analysis of the crude extract led to the identification of the quinone antibiotic γ-rubromycin, a promising antitumour and antibacterial antibiotic. To the best of our knowledge, there is currently no report on the production of γ-rubromycin by S. iakyrus. Therefore, this study suggests S. iakyrus TA 36 as the first-reported source of this unique bioactive secondary metabolite.
Assuntos
Quinonas , Streptomyces , Filogenia , RNA Ribossômico 16S/genética , Quinonas/farmacologia , Antibacterianos/farmacologiaRESUMO
The study aimed to assess the antifungal activity of twenty-five essential oils (EOs) and the potential synergistic activity of the most effective EOs against significant indoor fungi of the genus Aspergillus [A. fumigatus (KBio-122), A. flavus (KBio-134), A. terreus (KBio-145) and A. niger (KBio-202)]. The chemical composition of all EOs was evaluated by the gas chromatography coupled with mass spectrometry (GC/MS) and gas chromatography with flame ionization detector (GC-FID) analysis. The antifungal susceptibility of EOs was evaluated by using the broth microdilution method. The most effective EOs were selected to determine the minimum inhibitory concentrations (MICs) and minimum fungicidal concentrations (MFCs) at a concentration range from 256 to 0.125 µg/mL. For the synergistic activities, the most effective EOs were tested using the chessboard pattern. The most sensitive strain to treatments with essential oils alone and in the combination of EOs was A. flavus (KBio-134). The chessboard assay showed that combinations of lemongrass and thyme EOs proved the most potent synergistic antifungal activity (FICI = 0.1875) against A. fumigatus (KBio-122). The synergy displayed by a combination of some EOs may be used to control fungal growth or increasing resistance to available synthetic antifungals, consequently permitting the reduction of their most active doses.
Assuntos
Óleos Voláteis , Antifúngicos/farmacologia , Aspergillus , Fungos , Cromatografia Gasosa-Espectrometria de Massas , Testes de Sensibilidade Microbiana , Óleos Voláteis/farmacologiaRESUMO
Effective and reliable methods of identification of Pseudomonas species are important for the characterization of microorganisms. Freshwater ecosystems are an important source of Pseudomonas species, including those pathogenic to fish and humans. The aim of the present study was to compare the identification conducted with MALDI-TOF MS Biotyper and 16S rDNA sequencing of fish-borne Pseudomonas spp. Altogether, 13 different Pseudomonas spp. were isolated from freshwater fish. Phylogenetic analysis showed a clear taxonomic placement only for 13 out of 15 Pseudomonas isolates. Accordance of identification method was found only in 6 out of 15 isolates. The human pathogenic Pseudomonas spp. were not found in our study, indicating that the fish could be considered as safe for consumption. The present study revealed a high discriminatory power of the mass spectra investigation and 16S rDNA gene sequencing technology for the identification of Pseudomonas spp. associated with freshwater fish.
Assuntos
DNA Ribossômico/genética , Peixes/microbiologia , Pseudomonas/genética , Pseudomonas/isolamento & purificação , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz/métodos , Animais , Técnicas de Tipagem Bacteriana/métodos , DNA Bacteriano/genética , Filogenia , Pseudomonas/classificação , RNA Ribossômico 16S/genética , Análise de Sequência de DNA/métodosRESUMO
The main objective of this study was using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) for assembling of DSM (German Collection of Microorganisms) Streptomyces spectral database and identification of wild Streptomyces cultures, which were clustered by MALDI-TOF Biotyper OC software as well as for teracycline detection by observing of obtained spectra using flexAnalysis software. Production of tetracycline was confirmed by thin-layer chromatography. Presence of tetracycline mass spectrum was verified by several tetracycline producers (Streptomyces aureofaciens LMG 5968, S. aureofaciens 84/25, and S. aureofaciens BMK) and by pure tetracycline mass. Our results showed that it is possible to use MALDI-TOF MS for identification of tetracycline producers within Streptomyces genera by several easy steps. The purpose of this study was to establish cheap and quick detection of tetracycline producers.
Assuntos
Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz , Streptomyces/isolamento & purificação , Streptomyces/metabolismo , Tetraciclina/metabolismo , Bases de Dados Factuais , Ensaios de Triagem em Larga Escala/métodos , Humanos , Software , Tetraciclina/química , Tetraciclina/isolamento & purificaçãoRESUMO
The phylogenetic structure of 109 soil-borne entomopathogenic Beauveria isolates acquired using the Galleria mellonella bait method from different habitat types in Slovakia was determined by sequence analysis of their ITS and Bloc loci. Three Beauveria species were identified; Beauveria bassiana, B. pseudobassiana and B. brongniartii, represented by 51.4%, 43.1% and 5.5% of acquired isolates, respectively, which were resolved into 15, 1 and 1 distinguishable haplotypes. Correlation analysis with the habitat type and individual habitat characteristics showed strong preferences of the most prevalent haplotypes for agricultural (B. bassiana A1) and forest habitats (B. pseudobassiana) which has possible implications for conservative biocontrol strategies.
Assuntos
Beauveria/genética , Filogenia , Microbiologia do Solo , Animais , EslováquiaRESUMO
The aim of the present study was to assess the antifungal and anti-toxinogenic activity of 15 essential oils (EOs) against three fungi of the genus Aspergillus (A. parasiticus KMi-227-LR, A. parasiticus KMi-220-LR and A. flavus KMi-202-LR). The minimum inhibitory doses (MIDs) of the tested essential oils and their antifungal activity were determined using the micro-atmosphere method. The original commercial essential oil samples of Jasminum officinale L., Thymus vulgaris L., Syzygium aromaticum (L.) Merrill & Perry, Rosmarinus officinalis L., Ocimum basilicum L., Eucalyptus globulus Labill., Salvia officinalis L., Citrus limon (L.) Burm, Origanum vulgare L., Lavandula angustifolia Mill., Carum carvi L., Citrus sinensis (L.) Osbeck., Zingiber officinalis Rosc., Mentha piperita L. and Cinnamomum zeylanicum Nees. (C. verum J.S.Presl.) were produced in Slovakia (Calendula a.s., Nová Lubovna, Slovakia). All essential oils exhibited activity against all tested strains of fungi. After 14 days of incubation, A. flavus (KMi-202-LR) showed the highest susceptibility with a growth inhibition percentage (GIP) of 18.70% to C. limon and 5.92% to C. sinensis, while A. parasiticus (KMi-220-LR) exhibited a GIP of 20.56% to J. officinale. The minimum inhibitory doses (MIDs) of EOs with the most significant activity were recorded. The best antifungal activity, using the micro-atmosphere method was found in S. aromaticum with an MID of 62.5 µL L-1 air, T. vulgaris (MID of 62.5 µL L-1 air) and O. vulgare (MID of 31.5 µL L-1 air) against all tested strains. Mycotoxin production of the tested strains was evaluated by the thin layer chromatography (TLC) method. Mycotoxin production of AFB1 and AFG1 was inhibited following all treatments with C. carvi, R. officinale and S. officinale, Eucalyptus globulus L. and O. basilicum L. Essential oils exhibited a potential inhibition activity against toxic fungi, although, these affected only the production of AFB1.
Assuntos
Antifúngicos/efeitos adversos , Aspergillus/efeitos dos fármacos , Fungos/efeitos dos fármacos , Micotoxinas/biossíntese , Óleos Voláteis/farmacologia , Óleos de Plantas/farmacologia , Testes de Sensibilidade Microbiana , Óleos Voláteis/química , Óleos de Plantas/química , EslováquiaRESUMO
This investigation was undertaken to determine the impact of the insecticides Dursban 480 EC (with organophosphate compound chlorpyrifos as the active ingredient) and Talstar 10 EC (with pyrethroid bifenthrin as the active ingredient) on the respiration activity and microbial diversity in a sandy loam luvisol soil. The insecticides were applied in two doses: the maximum recommended dose for field application (15 mg kg(-1) for Dursban 480 EC and 6 mg kg(-1) for Talstar 10 EC) and a 100-fold higher dose for extrapolation of their effect. Bacterial and fungal genetic diversity was analysed in soil samples using PCR DGGE and the functional diversity (catabolic potential) was studied using BIOLOG EcoPlates at 1, 3, 7, 14, 28, 56 and 112 days after insecticide application. Five bacterial groups (α, ß, γ proteobacteria, firmibacteria and actinomycetes) and five groups of fungi or fungus-like microorganisms (Ascomycota, Basidiomycota, Chytridiomycota, Oomycota and Zygomycota) were analysed using specific primer sets. This approach provides high resolution of the analysis covering majority of microorganisms in the soil. Only the high-dose Dursban 480 EC significantly changed the community of microorganisms. We observed its negative effect on α- and γ-proteobacteria, as the number of OTUs (operational taxonomic units) decreased until the end of incubation. In the ß-proteobacteria group, initial increase of OTUs was followed by strong decrease. Diversity in the firmibacteria, actinomycetes and Zygomycota groups was minimally disturbed by the insecticide application. Dursban 480 EC, however, both positively and negatively affected certain species. Among negatively affected species Sphingomonas, Flavobacterium or Penicillium were detected, but Achromobacter, Luteibacter or Aspergillus were supported by applied insecticide. The analysis of BIOLOG plates using AWCD values indicated a significant increase in metabolic potential of microorganisms in the soil after the high-dose Dursban application. Analysis of respiration demonstrated high microbial activity after insecticide treatments; thus, microbial degradation was relatively fast. The half-life of the active insecticide compounds were estimated within the range of 25 to 27 days for Talstar and 6 to 11 days for Dursban and higher doses stimulated degradation. The recommended dose levels of both insecticides can be considered as safe for microbial community in the soil.
Assuntos
Bactérias/efeitos dos fármacos , Fungos/efeitos dos fármacos , Inseticidas/toxicidade , Oomicetos/efeitos dos fármacos , Microbiologia do Solo , Clorpirifos/toxicidade , Piretrinas/toxicidadeRESUMO
A total of 939 isolates of 11 genera representing 15 species of keratinophilic fungi were isolated and identified from the soils of three long-term fold-grazed pastures in national parks of Slovakia (Pod Ploskou, Strungový príslop, and Pod Keckou) and one non-fold-grazed pasture in sierra Stolicke vrchy (Diel) using the hair-baiting technique. Keratinophilic fungi were present in all soil samples with a prevalence of Trichophyton ajelloi and Paecilomyces lilacinus. These fungi were more abundant in soil from fold-grazed pasture (Strungový príslop) compared to non-fold-grazed pasture (Diel). The occurrence of the other keratinophilic fungi was substantially lower, likely because of low pH in some soils.
Assuntos
Biodiversidade , Fungos/isolamento & purificação , Fungos/metabolismo , Queratinas/metabolismo , Microbiologia do Solo , Fungos/classificação , Fungos/crescimento & desenvolvimento , Cabelo/microbiologia , EslováquiaRESUMO
Diverse communities of bacterial endophytes inhabit plant tissues, and these bacteria play important roles for plant growth and health. Cherry laurel (Prunus laurocerasus L.) is a broadleaf evergreen shrub that is widely grown in temperate zones for its ornamental and medicinal properties, however virtually nothing is known about its associated bacterial community. In this study, we analysed the matured one-year-old leaves of this plant using Illumina-based 16S rRNA gene metabarcoding to reveal the community structure of endophytic bacteria and understand its shifts during the seasonal transition from winter dormancy to a spring vegetative state. The overall community was composed of four dominant phyla (Proteobacteria, Actinobacteria, Firmicutes, Bacteroidetes). Corynebacterium, Acinetobacter, and Chryseobacterium genera were the most prevalent bacteria, comprising 13.3%, 6.9%, and 6.8% of the amplicon sequence variants (ASVs), respectively. The ASV richness and diversity increased significantly in May as compared to other sampling months (February, March, and April). We observed high variation in the overall community structure of endophytic bacteria among collection dates. The variation was only reflected by a few core community members, suggesting that the changes of the endophytic community during winter/spring seasonal transition are mostly associated with the less abundant community members. We identified biomarker taxa for late winter, mid spring, and late spring collection dates. This study is the first one to report on the diversity and composition of bacterial endophytes in the leaves of cherry laurel and its shifts across the dormancy-to-vegetative seasonal transition.
RESUMO
This study aimed to determine the in vitro and in situ antifungal activity of (14) selected essential oils (EOS), namely clove, thyme, red thyme, litsea, eucalyptus, niaouli, fennel, anise, cumin, basil, rosemary, sage, bergamot mint, and marjoram, by vapor contact against the growth of two strains of Penicillium commune (KMi-183 and KMi-402). Furthermore, to exclude the negative effect of EOs on the lactic acid bacteria (LABs) (Streptococcus spp.) on cheeses, their influence was monitored. Next, the sensory evaluation of cheese treated by EOs was evaluated. The results show that litsea and clove EOs were the most effective in the vapor phase against both tested strains. These EOs were characterized by the highest amount of α- (40.00%) and ß-Citral (34.35%) in litsea and eugenol (85.23%) in clove. The antitoxicogenic activity of less effective (in growth inhibition) EOs on cyclopiazonic acid (CPA) production by the tested strains was also observed. The growth of Streptococcus spp. (ranging from 8.11 to 9.69 log CFU/g) was not affected by the EOs in treated cheese. Even though the evaluators recognized some EOs in sensory evaluation by the triangle test, they did not have a negative effect on the taste and smell of the treated cheeses and were evaluated as edible. The antifungal activity of EOs against several types of microscopic fungi and their effect on the sensory properties of treated foods needs to be further tested to achieve the most effective protection of foods from their direct contaminants.
RESUMO
Essential oils (EOs) from aromatic plants seem to have the potential to control several fungal pathogens and food contaminants. Botrytis cinerea is the main strawberry fruit contaminant causing high losses during storage. Here, thirteen EOs applied in the vapor phase were evaluated for their potential to inhibit the growth of three different strains of B. cinerea isolated from strawberry fruits. Eight EOs (lemongrass, litsea, lavender, peppermint, mint, petitgrain, sage, and thyme) were able to completely inhibit the growth of B. cinerea for 7 days when applied at a concentration of 625 µL·L-1. Four EOs with the lowest minimal inhibition concentrations (thyme, peppermint, lemongrass, and litsea) have been tested on strawberry fruits intentionally inoculated by B. cinerea. All four EOs showed high inhibition at a concentration of 250 or 500 µL·L-1, but only peppermint EO was able to completely inhibit B. cinerea lesion development at a concentration of 125 µL·L-1. The sensory evaluation of strawberries treated by EOs at a concentration 125 µL·L-1 resulted in a statistically significant decrease in taste, aftertaste, aroma, and overall quality. Lemongrass and litsea EOs scored better than thyme and peppermint ones, thus forming two viable methods for B. cinerea suppression and the extension of packed strawberries' shelf life.
RESUMO
Klebsiella pneumoniae carbapenemase (KPC)-producing bacteria is a group of highly dangerous antibiotic resistant Gram-negative Enterobacteriaceae. They cause infections associated with significant morbidity and mortality. Therefore, the rapid detection of KPC-producing bacteria plays a key role in clinical microbiology. Matrix assisted laser desorption/ionization time-of- flight (MALDI-TOF) is a rapidly evolving technology that finds application in various clinical, scientific, and industrial disciplines. In the present study, we demonstrated three different procedures of carbapenemase-producing K. pneumoniae (KPC) detection. The most basic model of MALDI-TOF instrument MS Microflex LT was used, operating in the linear ion-positive mode, commonly used in modern clinical laboratories. The first procedure was based on indirect monitoring of carbapenemase production with direct detection of hydrolyzed carbapenem antibiotic degradation products in the mass spectrum. The second procedure was based on direct detection of blaKPC accompanying peak with an 11,109 Da in the mass spectrum of carbapenemase-producing K. pneumoniae (KPC), which represents the cleaved protein (pKpQIL_p019) expressed by pKpQIL plasmid. In addition, several unique peaks were detected in the carbapenemase-producing K. pneumoniae (KPC) mass spectrum. The third procedure was the identification of carbapenemase-producing K. pneumoniae (KPC) based on the protein fingerprint using local database created from the whole mass spectra. By comparing detection procedures, we determined that the third procedure was very fast and relatively easy. However, it requires previous verification of carbapenemase-producing K. pneumoniae (KPC) using other methods as genetic blaKPC identification, detection of carbapenem degradation products, and accompanying peak with 11,109 Da, which represents cleaved pKpQIL_p019 protein expressed by pKpQIL plasmid. Detection of carbapenemase-producing K. pneumoniae using MALDI-TOF provides fast and accurate results that may help to reduce morbidity and mortality in hospital setting when applied in diagnostic situations.
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Soil microorganisms and their activities are essential for maintaining soil health and fertility. Microorganisms can be negatively affected by application of herbicides. Although effects of herbicides on microorganisms are widely studied, there is a lack of information for chloroacetamide herbicide dimethachlor. Thus, dimethachlor and well known linuron were applied to silty-loam luvisol and their effects on microorganisms were evaluated during112 days long laboratory assay. Dimethachlor and linuron were applied in doses 1.0 kg ha-1 and 0.8 kg ha-1 corresponding to 3.33 mg kg-1 and 2.66 mg kg-1 respectively. Also 100-fold doses were used for magnification of impacts. Linuron in 100-fold dose caused minor increase of respiration, temporal increase of soil microbial biomass, decrease of soil dehydrogenase activity, and altered microbial community. Dimethachlor in 100-fold dose significantly increased respiration; microbial biomass and decreased soil enzymatic activities. Microbial composition changed significantly, Proteobacteria abundance, particularly Pseudomonas and Achromobacter genera increased from 7 to 28th day. In-silico prediction of microbial gene expression by PICRUSt2 software revealed increased expression of genes related to xenobiotic degradation pathways. Evaluated characteristics of microbial community and activity were not affected by herbicides in recommended doses and the responsible use of both herbicides will not harm soil microbial community.
Assuntos
Acetamidas/toxicidade , Linurona/toxicidade , Microbiota/efeitos dos fármacos , Microbiologia do Solo , Aerobiose/efeitos dos fármacos , Biomassa , Dióxido de Carbono/metabolismo , Herbicidas/toxicidade , Redes e Vias Metabólicas/efeitos dos fármacos , FilogeniaRESUMO
Bacterial contamination of semen is an important factor connected to the health status of bulls that may significantly affect semen quality for artificial insemination. Moreover, some important bovine diseases may be transmitted through semen. Up to now, only a very limited number of complex studies describing the semen microbiome of bulls have been published, as many bacteria are hard to cultivate using traditional techniques. The 16S rRNA high-throughput sequencing strategy allows for the reliable identification of bacterial profiles of bovine semen together with the detection of noncultivable bacterial species. Fresh samples from Holstein Friesian breeding bulls (n = 55) were examined for the natural variability in the present bacteria. Semen doses were selected randomly from Slovak Biological Services in Nitra, Slovak Republic. The most predominant phyla within the whole dataset were Firmicutes (31%), Proteobacteria (22%), Fusobacteria (18%), Actinobacteria (13%) and Bacteroidetes (12%). Samples of semen were divided into two separate clusters according to their microbiome compositions using a cording partition around a medoids analysis. Microbiomes of the first cluster (CL1) of samples (n = 20) were based on Actinobacteria (CL1 average = 25%; CL = 28%) and Firmicutes (CL1 = 38%; CL2 = 27%), while the second cluster (CL2; n = 35) contained samples characterized by a high prevalence of Fusobacteria (CL1 = 4%; CL2 = 26%). Some important indicator microbial groups were differentially distributed between the clusters.
RESUMO
The main objective of this study is to evaluate the effect of selected essential oils thyme chemotype linalool (Thymus zygis L.), thyme chemotype tymol (Thymus vulgaris L.), eucalyptus (Eucalyptus globulus Labill.), lavender (Lavandula angustifolia Mill.), mint (Mentha piperita L.), almond (Prunbus dulcis Mill.), cinnamon bark (Cinnamomum zeylanicum Nees), litsea (Litsea cubeba Lour. Pers), lemongrass (Cympogon citrati L. Stapf), and ginger (Zingiber officinalis Rosc.) in the vapor phase on growth, sporulation, and mycotoxins production of two Aspergillus strains (Aspergillus parasiticus CGC34 and Aspergillus ochraceus CGC87), important postharvest pathogens of green and roasted coffee beans. Moreover, the effect of the essential oils (EOs) on the sensory profile of the coffee samples treated with EOs was evaluated. The major components of tested EOs were determined by gas chromatography and mass spectrometry (GC-MS) and gas chromatography with flame ionization detector (GC-FID). The results showed that almond, cinnamon bark, lemongrass, and litsea EOs are able to significantly inhibit the growth, sporulation, and mycotoxins production by toxigenic fungi. Sensory evaluation of coffee beans treated with EOs before and after roasting showed that some EOs (except lemongrass and litsea) do not adversely affect the taste and aroma of coffee beverages. Thus, application of the vapors of almond and cinnamon EOs appears to be an effective way that could serve to protect coffee during its transport and storage from toxigenic fungi.
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Endophytic bacteria are an important part of different functions in plants that lead to plants' production characteristics as well as their stress response mechanisms. Endophytic bacterial diversity was analyzed in this study to describe 16S rRNA variability and changes in the leaves of drought-tolerant and drought-susceptible wheat when growth under in vitro conditions. A metagenomic analysis was applied and a pilot exploratory study was performed to prove this type of analysis as applicable to tracking endophytic bacterial diversity changes when a drought stress is applied to an in vitro culture of wheat. The study showed that the changes in the bacterial endophytes' variabilities associated preferentially with the drought stress varietal characteristics of the analyzed wheat instead of the applied stress conditions.
RESUMO
Sulfonylurea herbicides are widely used for weed control in agriculture, and they are suspected to alter microbial communities and activities in the soil. This study investigates the impact of two sulfonylurea herbicides chlorsulfuron and sulfosulfuron on microbial community and activity in two different soils taken from two sites in west part of the Slovak Republic. The soil from the Malanta site was silt-loam luvisol with pH(H2O) 5.78 while the soil from the Stefanov site was sandy-loam regosol with pH(H2O) 8.25. These soils were not treated by sulfonylurea herbicides at least for 2 years prior to the study. In laboratory assay, the herbicides were applied to soil in their maximal recommended doses 26 and 25 g per hectare of chlorsulfuron and sulfosulfuron, respectively. Their effect was evaluated on the 3rd, 7th, 14th, 28th, 56th, and 112th day after application to soil. Illumina high-throughput amplicon sequencing of the 16S rRNA gene and ITS region was used to monitor changes on prokaryotic and fungal community composition. Enzymatic activity was evaluated using 11 substrates. Physiological profile of microbial community was analyzed using Biolog© ecoplates. Significant changes in enzymatic activity caused by the application of herbicides were found during the first 28 days. The application of herbicides altered the activity of cellobiohydrolase, arylsulphatase, dehydrogenase, phosphatase, and FDA hydrolase. Chlorsulfuron caused a more varying response of enzymatic activity than sulfosulfuron, and observed changes were not the same for both soils. In Malanta soil, chlorsulfuron decreased dehydrogenase activity while it was increased in the Stefanov soil. Phosphatase activity was decreased in both soils on 7th and 14th day. There were only minor changes in prokaryotic or fungal community or physiological profiles regarding pesticide application. Differences between soils and incubation time explained most of the variability in these parameters. Diversity indices, physiological parameters, and enzymatic activity decreased over time. The results have shown that chlorsulfuron and sulfosulfuron can affect the function and activity of the soil microbial community without significant change in its composition.
Assuntos
Herbicidas , Microbiota , Poluentes do Solo , Agricultura , Herbicidas/análise , Pirimidinas , RNA Ribossômico 16S , Eslováquia , Solo , Microbiologia do Solo , Poluentes do Solo/análise , Sulfonamidas , TriazinasRESUMO
Coconut oil has a high content of lauric acid, which has selective antibacterial activity. This study aimed to explore the effect of coconut oil ingestion on the gastrointestinal microbiomes of pigs. A 14-day-long feeding experiment included 19 pigs in two groups (9 on a normal diet and 10 on a diet supplemented with coconut oil). At the start and end of the experiment, a rectal swab sample was taken from each pig in both groups, and total bacterial DNA was extracted. We used 16S rRNA high-throughput amplicon sequencing to evaluate the microbiome changes during the feeding experiment. A total of 446 operational taxonomic units (OTUs) were detected in the whole sample set. Shannon's indices of bacterial diversity did not change significantly during the experiment. Changes in the bacterial community during the study period and in response to the coconut oil treatment were highly significant (p < 0.001). During the study, an increase in the abundance of Lactobacillus was detected in the group treated with coconut oil. An increase in Alloprevotella, Bifidobacteriales, and Lactobacillales and a decrease in Corynebacterium, Mitsuokella, Psychrobacter, and Pseudomonadales were attributed to the coconut oil treatment. Although the addition of coconut oil to pig feed did not affect Shannon's index of diversity, it had a positive effect on the abundance of bacterial groups that are considered to be commensal and/or probiotic.
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Fifty seven soil-borne actinomycete strains were assessed for the antibiotic production. Two of the most active isolates, designed as Streptomyces ST-13 and DK-15 exhibited a broad range of antimicrobial activity and therefore they were selected for HPLC fractionation against the most suppressed bacteria Staphylococcus aureus (ST-13) and Chromobacterium violaceum (DK-15). LC/MS analysis of extracts showed the presence of polyketides factumycin (DK15) and tetrangomycin (ST13). The taxonomic position of the antibiotic-producing actinomycetes was determined using a polyphasic approach. Phenotypic characterization and 16S rRNA gene sequence analysis of the isolates matched those described for members of the genus Streptomyces. DK-15 strain exhibited the highest 16S rRNA gene sequence similarity to Streptomyces globosus DSM-40815 (T) and Streptomyces toxytricini DSM-40178 (T) and ST-13 strain to Streptomyces ederensis DSM-40741 (T) and Streptomyces phaeochromogenes DSM-40073 (T). For the proper identification, MALDI-TOF/MS profile of whole-cell proteins led to the identification of S. globosus DK-15 (accession number: KX527570) and S. ederensis ST13 (accession number: KX527568). To our knowledge, there is no report about the production of these antibiotics by S.globosus and S. ederensis, thus isolates DK15 and ST13 identified as S. globosus DK-15 and S.ederensis ST-13 can be considered as new sources of these unique antibacterial metabolites.
Assuntos
Antibacterianos/biossíntese , Streptomyces/isolamento & purificação , Streptomyces/metabolismo , Técnicas de Tipagem Bacteriana , Benzo(a)Antracenos/metabolismo , DNA Bacteriano/genética , Filogenia , Piridonas/metabolismo , Microbiologia do Solo , Streptomyces/classificação , Streptomyces/genéticaRESUMO
Microbiological investigation of unexplored ecosystems is crucial for discovering of antibiotic producing actinomycetes. The present study was conducted to determine antimicrobial activity and identify the most active strains. Actinomycetes were isolated using the spread plate technique following by serial dilution of samples on starch casein agar. The screening method consists of primary and secondary testing. The most active isolates were identified based on molecular and cultural methods. 42 out of 66 isolates displayed antimicrobial potential. 63% exhibited antibacterial activity, 16% antifungal activity, and 16% displayed both activities. Identified isolates, Streptomyces scabrisporus, Streptomyces sparsogenes, Streptomyces misakiensis, Streptomyces cirratus, Streptomyces lincolnensis, Streptomyces endophyticus, Streptomyces chartreusis, and Streptomyces alboniger showed a broad spectrum of enzymatic activities. The results indicated that these isolates may serve as antibiotic and enzyme-producing microbes.