RESUMO
Ruminants are considered to be less sensitive towards mycotoxins than monogastric animals because rumen microbiota have mycotoxin-detoxifying capacities. Therefore the effect of mycotoxins towards ruminants has been studied to a lesser extent compared with monogastric animals. Worldwide, a high proportion of the ruminant diet consists of silages made of forage crops (i.e. all parts of the crop above the stubble are harvested). In practice, silages are often contaminated with multiple mycotoxins. Exposure to a cocktail of mycotoxins can hamper animal production and have severe health consequences. In this article the different aspects associated with mycotoxin contamination of silage are reviewed 'from seed to feed'. An overview is given on the occurrence of toxigenic fungal species and their concomitant mycotoxins in forage crops before and after ensiling. The mycotoxin load of visually non-mouldy samples and mouldy hot spots within the same silo is also compared. Subsequently, this review delves into different problem-solving strategies. A logical first step is prevention of mould growth and mycotoxin production in the field, during harvest and during ensiling. If prevention should fail, several remediation strategies are available. These are listed, mainly focusing on the possibilities of microbial degradation of mycotoxins in vivo in silage. © 2015 Society of Chemical Industry.
Assuntos
Contaminação de Alimentos/prevenção & controle , Fungos/classificação , Micotoxinas/química , Silagem/análise , Silagem/microbiologia , AnimaisRESUMO
Within sustainable resource management, the recovery of nitrogen and phosphorus nutrients from waste streams is becoming increasingly important. Although the use of microalgae has been described extensively in environmental biotechnology, the potential of nitrate-accumulating microalgae for nutrient recovery has not been investigated yet. The ability of these marine microorganisms to concentrate environmental nitrate within their biomass is remarkable. The aim of this study was to investigate the application potential of nitrate-accumulating diatoms for nutrient recovery from marine wastewaters. The intracellular nitrate storage capacity was quantified for six marine diatom strains in synthetic wastewater. Amphora coffeaeformis and Phaeodactylum tricornutum stored the highest amount of nitrate with respectively 3.15 and 2.10 g N L(-1) of cell volume, which accounted for 17.3 and 4.6 %, respectively, of the total nitrogen content. The growth and nitrate and phosphate uptake of both diatoms were further analyzed and based on these features P. tricornutum showed the highest potential for nutrient recovery. A mathematical model was developed which included intracellular nitrate storage and the kinetic parameters were derived for P. tricornutum. Furthermore, a simulation study was performed to compare the performance of a proposed microalgal nutrient recovery unit with a conventional denitrification system for marine wastewater treatment. Overall, this study demonstrates the potential application of P. tricornutum for saline wastewater treatment with concurrent nitrogen and phosphorus recycling.
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Diatomáceas/química , Diatomáceas/metabolismo , Microalgas/química , Microalgas/metabolismo , Nitratos/metabolismo , Biodegradação Ambiental , Diatomáceas/crescimento & desenvolvimento , Cinética , Microalgas/crescimento & desenvolvimento , Águas Residuárias/análiseRESUMO
Zearalenone (ZEN) is a prevalent mycotoxin found in grains and grain-derived products, inducing adverse health effects in both animals and humans. The in-field application of microorganisms to degrade and detoxify ZEN is a promising strategy to enhance the safety of food and feed. In this study, we investigated the potential of three actinobacterial strains to degrade and detoxify ZEN in vitro and in planta on wheat ears. The residual ZEN concentration and toxicity in the samples were analysed with UHPLC-MS/MS and a bioluminescence BLYES assay, respectively. Streptomyces rimosus subsp. rimosus LMG19352 could completely degrade and detoxify 5 mg/L ZEN in LB broth within 24 h, along with significant reductions in ZEN concentration both in a minimal medium (MM) and on wheat ears. Additionally, it was the only strain that showed a significant colonisation of these ears. Rhodococcus sp. R25614 exhibited partial but significant degradation in LB broth and MM, whereas Streptomyces sp. LMG16995 degraded and detoxified ZEN in LB broth after 72 h by 39% and 33%, respectively. Although all three actinobacterial strains demonstrated the metabolic capability to degrade and detoxify ZEN in vitro, only S. rimosus subsp. rimosus LMG19352 showed promising potential to mitigate ZEN in planta. This distinction underscores the importance of incorporating in planta screening assays for assessing the potential of mycotoxin-biotransforming microorganisms as biocontrol agents.
Assuntos
Agentes de Controle Biológico , Triticum , Zearalenona , Zearalenona/metabolismo , Zearalenona/toxicidade , Triticum/microbiologia , Agentes de Controle Biológico/metabolismo , Streptomyces/metabolismo , Actinobacteria/metabolismo , Contaminação de Alimentos/prevenção & controle , Espectrometria de Massas em TandemRESUMO
To investigate whether the pesticide removal in on-farm biopurification systems (BPS) filled with two different types of substrata (biomix and plastic carriers) is affected by inoculation with a pesticide-degrading strain or mixed culture, lab-scale BPS used to treat chloropropham point source contaminations were bioaugmented with either a specialized chloropropham-degrading strain or a chloropropham-degrading enrichment culture. Application of both inoculum types leads to an accelerated degradation activity in the columns filled with plastic carriers. For both substratum types, inoculation with the mixed culture resulted in a lower breakthrough of the toxic intermediate 3-chloroaniline at high hydraulic loads, compared to inoculation with the pure isolate and no inoculation. This study suggests that the use of plastic carrier materials could be a proficient alternative to the use of a conventional biomix as a substratum in on-farm BPS and that inoculation with a mixed degrading culture can reduce the leaching of more mobile toxic intermediates.
Assuntos
Inoculantes Agrícolas/metabolismo , Bactérias/metabolismo , Recuperação e Remediação Ambiental/métodos , Praguicidas/metabolismo , Poluentes Químicos da Água/metabolismo , Inoculantes Agrícolas/classificação , Inoculantes Agrícolas/genética , Bactérias/classificação , Bactérias/genética , Biodegradação Ambiental , Reatores Biológicos/microbiologia , Recuperação e Remediação Ambiental/instrumentação , Purificação da Água/métodosRESUMO
Pathogens resistant to multiple antibiotics are rapidly emerging, entailing important consequences for human health. This study investigated if the broad-host-range multiresistance plasmid pB10, isolated from a wastewater treatment plant, harbouring amoxicillin, streptomycin, sulfonamide, and tetracycline resistance genes, was transferable to the foodborne pathogens Salmonella spp. or E. coli O157:H7 and how this transfer alters the phenotype of the recipients. The transfer ratio was determined by both plating and flow cytometry. Antibiotic resistance profiles were determined for both recipients and transconjugants using the disk diffusion method. For 14 of the 15 recipient strains, transconjugants were detected. Based on plating, transfer ratios were between 6.8 × 10â»9 and 3.0 × 10⻲ while using flow cytometry, transfer ratios were between <1.0 × 10â»5 and 1.9 × 10⻲. With a few exceptions, the transconjugants showed phenotypically increased resistance, indicating that most of the transferred resistance genes were expressed. In summary, we showed that an environmental plasmid can be transferred into foodborne pathogenic bacteria at high transfer ratios. However, the transfer ratio seemed to be recipient strain dependent. Moreover, the newly acquired resistance genes could turn antibiotic susceptible strains into resistant ones, paving the way to compromise human health.
Assuntos
Resistência Microbiana a Medicamentos/genética , Escherichia coli O157/genética , Transferência Genética Horizontal , Plasmídeos/genética , Salmonella/genética , Antibacterianos/farmacologia , Infecções por Escherichia coli/microbiologia , Escherichia coli O157/efeitos dos fármacos , Citometria de Fluxo , Doenças Transmitidas por Alimentos/microbiologia , Humanos , Fenótipo , Reação em Cadeia da Polimerase , Salmonella/efeitos dos fármacos , Infecções por Salmonella/microbiologiaRESUMO
As the awareness on the ecological impact of chemical phosphate fertilizers grows, research turns to sustainable alternatives such as the implementation of phosphate solubilizing bacteria (PSB), which make largely immobile phosphorous reserves in soils available for uptake by plants. In this review, we introduce the mechanisms by which plants facilitate P-uptake and illustrate how PSB improve the bioavailability of this nutrient. Next, the effectiveness of PSB on increasing plant biomass and P-uptake is assessed using a meta-analysis approach. Our review demonstrates that improved P-uptake does not always translate in improved plant height and biomass. We show that the effect of PSB on plants does not provide an added benefit when using bacterial consortia compared to single strains. Moreover, the commonly reported species for P-solubilization, Bacillus spp. and Pseudomonas spp., are outperformed by the scarcely implemented Burkholderia spp. Despite the similar responses to PSB in monocots and eudicots, species responsiveness to PSB varies within both clades. Remarkably, the meta-analysis challenges the common belief that PSB are less effective under field conditions compared to greenhouse conditions. This review provides innovative insights and identifies key questions for future research on PSB to promote their implementation in agriculture.
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The use of phosphate solubilizing bacteria (PSB) as inoculants for the rhizosphere is a well-known strategy to mitigate P-deficiency in plants. However, despite the multiple modes of action to render P available for plants, PSB often fail to deliver in the field as their selection is often based on a single P-solubilizing trait assessed in vitro. Anticipating these shortcomings, we screened 250 isolates originating from rhizosphere-based enriched consortia for the main in vitro P-solubilizing traits, and subsequently grouped the isolates through trait-based HCPC (hierarchical clustering on principal components). Representative isolates of each cluster were tested in an in planta experiment to compare their in vitro P-solubilizing traits with their in planta performance under conditions of P-deprivation. Our data convincingly show that bacterial consortia capable to mitigate P-deficiency in planta were enriched in bacterial isolates that had multiple P-solubilizing traits in vitro and that had the capacity to mitigate plant P-stress in planta under P-deprived conditions. Furthermore, although it was assumed that bacteria that looked promising in vitro would also have a positive effect in planta, our data show that this was not always the case. Opposite, lack of performance in vitro did not automatically result in a lack of performance in planta. These results corroborate the strength of the previously described in planta-based enrichment and selection technique for the isolation of highly efficient rhizosphere competent PSB. IMPORTANCE With the growing awareness on the ecological impact of chemical phosphate fertilizers, research concerning the use of phosphate solubilizing bacteria (PSB) as a sustainable alternative for, or addition to these fertilizers is of paramount importance. In previous research, we successfully implemented a plant-based enrichment technique for PSB, which simultaneously selected for the rhizosphere competence and phosphate solubilizing characteristics of bacterial suspensions. Current research follows up on our previous findings, whereas we screened 250 rhizobacteria for their P-solubilizing traits and were able to substantiate the results obtained from the enriched suspensions at a single-isolate level. With this research, we aim for a paradigm shift toward the plant-based selection of PSB, which is a more holistic approach compared to the plate-based methods. We emphasize the strength of the previously described plant-based enrichment and selection technique for the isolation of highly efficient and diverse PSB.
Assuntos
Fertilizantes , Rizosfera , Fosfatos , Bactérias , Microbiologia do SoloRESUMO
This work introduces an alternative workflow for the discovery of novel bacterial biocontrol agents in wheat against Fusarium head blight (FHB). Unlike the mass testing of isolate collections, we started from a diverse inoculum by extracting microbiomes from ears of field-grown plants at grain filling stage. Four distinct microbial communities were generated which were exposed to 3 14-day culture-independent experimental enrichments on detached wheat spikes infected with F. graminearum PH1. We found that one bacterial community reduced infection symptoms after 3 cycles, which was chosen to subsequently isolate bacteria through limiting dilution. All 94 isolates were tested in an in vitro and in planta assay, and a selection of 14 isolates was further tested on detached ears. The results seem to indicate that our enrichment approach resulted in bacteria with different modes-of-action in regard to FHB control. Erwinia persicina isolate C3 showed a significant reduction in disease severity (Fv/Fm), and Erwinia persicina C3 and Pseudomonas sp. B3 showed a significant reduction in fungal biomass (cGFP). However, the mycotoxin analysis of both these treatments showed no reduction in DON levels. Nevertheless, Pantoea ananatis H3 and H11 and Erwinia persicina H2 were able to reduce DON concentrations by more than 50%, although these effects were not statistically significant. Lastly, Erwinia persicina H2 also showed a significantly greater glucosylation of DON to the less phytotoxic DON-3G. The bacterial genera isolated through the enrichment cycles have been reported to dominate microbial communities that develop in open habitats, showing strong indications that the isolated bacteria can reduce the infection pressure of F. graminearum on the spike phyllosphere.
Assuntos
Erwinia , Fusarium , Tricotecenos , Doenças das Plantas/microbiologia , Doenças das Plantas/prevenção & controle , Triticum/microbiologiaRESUMO
Chloropropham-degrading cultures were obtained from sludge and soil samples by using two different enrichment techniques: (i) planktonic enrichments in shaken liquid medium and (ii) biofilm enrichments on two types of solid matrixes (plastic chips and gravel). Denaturing gradient gel electrophoresis fingerprinting showed that planktonic and biofilm cultures had a different community composition depending on the presence and type of added solid matrix during enrichment. This was reflected in the unique chloropropham-degrading species that could be isolated from the different cultures. Planktonic and biofilm cultures also differed in chloropropham-degrading activity. With biofilm cultures, slower chloropropham removal was observed, but with less build-up of the toxic intermediate 3-chloroaniline. Disruption of the biofilm architecture resulted in degradation characteristics shifting toward those of the free suspensions, indicating the importance of a well-established biofilm structure for good performance. These results show that biofilm-mediated enrichment techniques can be used to select for pollutant-degrading microorganisms that like to proliferate in a biofilm and that cannot be isolated using conventional shaken-liquid procedures. Furthermore, the influence of the biofilm architecture on the pesticide degradation characteristics suggests that for bioaugmentation the use of biofilm catabolic communities might be a proficient alternative to using planktonic freely suspended cultures.
Assuntos
Adaptação Fisiológica/efeitos dos fármacos , Bactérias/efeitos dos fármacos , Biofilmes/efeitos dos fármacos , Praguicidas/metabolismo , Plâncton/efeitos dos fármacos , Compostos de Anilina , Bactérias/isolamento & purificação , Bactérias/metabolismo , Biodegradação Ambiental , Clorprofam/análise , Clorprofam/química , Clorprofam/metabolismo , Meios de Cultura , Mapeamento de Peptídeos , Praguicidas/farmacologia , Plâncton/metabolismo , RNA Ribossômico 16S/metabolismo , Esgotos/microbiologia , Solo , Microbiologia do SoloRESUMO
Phosphorus (P) is despite its omnipresence in soils often unavailable for plants. Rhizobacteria able to solubilize P are therefore crucial to avoid P deficiency. Selection for phosphate-solubilizing bacteria (PSB) is frequently done in vitro; however, rhizosphere competence is herein overlooked. Therefore, we developed an in planta enrichment concept enabling simultaneous microbial selection for P-solubilization and rhizosphere competence. We used an ecologically relevant combination of iron- and aluminium phosphate to select for PSB in maize (Zea mays L.). In each consecutive enrichment, plant roots were inoculated with rhizobacterial suspensions from plants that had grown in substrate with insoluble P. To assess the plants' P statuses, non-destructive multispectral imaging was used for quantifying anthocyanins, a proxy for maize's P status. After the third consecutive enrichment, plants supplied with insoluble P and inoculated with rhizobacterial suspensions showed a P status similar to plants supplied with soluble P. A parallel metabarcoding approach uncovered that the improved P status in the third enrichment coincided with a shift in the rhizobiome towards bacteria with plant growth-promoting and P-solubilizing capacities. Finally, further consecutive enrichment led to a functional relapse hallmarked by plants with a low P status and a second shift in the rhizobiome at the level of Azospirillaceae and Rhizobiaceae.
Assuntos
Microbiologia do Solo , Zea mays , Antocianinas , Bactérias/genética , Fosfatos , Raízes de PlantasRESUMO
Deoxynivalenol (DON) is a type B trichothecene mycotoxin with worldwide high incidence in feed which is produced by Fusarium species. Strategies are needed to eliminate its health risk for livestock and to minimise its economic impact on production. In order to assess the efficacy of potential physical, chemical and biological DON detoxifying agents, a good in vitro model is necessary to perform a fast and high-throughput screening of new compounds before in vivo trials are set up. In this paper, an in vitro model was developed to screen potential commercial products for DON degradation and detoxification. Contaminated feed with potential detoxifying agents are first applied to a simulated gastrointestinal tract (GIT) of a pig, after which detoxification is assessed through a robust, inexpensive and readily applicable Lemna minor L. aquatic plant bioassay which enables evaluation of the residual toxicity of possible metabolites formed by DON detoxifying agents. The GIT simulation enables taking matrix and incubation parameters into account as they can affect the binding, removal or degradation of DON. One product could reduce DON in feed in the GIT model for almost 100% after 6 h. DON metabolites were tentatively identified with LC-MS/MS. This GIT simulation coupled to a detoxification bioassay is a valuable model for in vitro screening and assessing compounds for DON detoxification, and could be expanded towards other mycotoxins.
Assuntos
Ração Animal/análise , Trato Gastrointestinal/metabolismo , Tricotecenos/análise , Tricotecenos/metabolismo , Animais , Cromatografia Líquida de Alta Pressão , Grão Comestível/microbiologia , Escherichia coli/metabolismo , Contaminação de Alimentos , Fusarium/metabolismo , Ensaios de Triagem em Larga Escala , Técnicas In Vitro , Lactobacillus acidophilus/metabolismo , Lactococcus lactis/metabolismo , Modelos Animais , Desintoxicação por Sorção , Suínos , Espectrometria de Massas em Tandem , Fatores de TempoRESUMO
The mycotoxin deoxynivalenol (DON), produced in wheat, barley and maize by Fusarium graminearum and Fusarium culmorum, is threatening the health of humans and animals. With its worldwide high incidence in food and feed, mitigation strategies are needed to detoxify DON, maintaining the nutritional value and palatability of decontaminated commodities. A promising technique is biological degradation, where microorganisms are used to biotransform mycotoxins into less toxic metabolites. In this study, bacterial enrichment cultures were screened for their DON detoxification potential, where DON and its potential derivatives were monitored. The residual phytotoxicity was determined through a bioassay using the aquatic plant Lemna minor L. Two bacterial enrichment cultures were found to biotransform DON into a still highly toxic metabolite for plants. Furthermore, a cytotoxic effect was observed on the cellular viability of intestinal porcine epithelial cells. Through liquid chromatography high-resolution mass spectrometry analysis, an unknown compound was detected, and tentatively characterized with a molecular weight of 30.0 Da (i.e., CH2O) higher than DON. Metabarcoding of the subsequently enriched bacterial communities revealed a shift towards the genera Sphingopyxis, Pseudoxanthomonas, Ochrobactrum and Pseudarthrobacter. This work describes the discovery of a novel bacterial DON-derived metabolite, toxic to plant and porcine cells.
Assuntos
Bactérias/metabolismo , Tricotecenos/metabolismo , Animais , Araceae/efeitos dos fármacos , Bactérias/genética , Técnicas Bacteriológicas , Biotransformação , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Código de Barras de DNA Taxonômico , Células Epiteliais/efeitos dos fármacos , Suínos , Tricotecenos/toxicidadeRESUMO
Due to the beneficial properties of silver, it is anticipated that the number of commercially available applications will keep growing during the next decade. In this study, 14 different commercial products that claim to contain solid silver were characterized by visual analysis, UV-VIS spectroscopy, inductive coupled plasma optical emission spectrometry (ICP-OES), scanning transmission electron microscopy with energy dispersive x-ray spectroscopy (STEM-EDX), and dynamic light scattering (DLS). Moreover the variation between production batches-which has never been researched before-was investigated. All four techniques corroborated that some products were highly concentrated and contained spherically-shaped silver nanoparticles (AgNPs), while in others, no (solid) silver was detected or only irregularly-shaped silver particles with a high size polydispersity were present. For almost all products, a significant difference between the claimed and measured silver concentration was detected and a high variability between different production batches of the same product was observed. Our results show the need for a more rigorous approach regarding the manufacturing, labeling, and use of silver-containing products.
RESUMO
Little is known about the range of hosts in which broad-host-range (BHR) plasmids can persist in the absence of selection for plasmid-encoded traits, and whether this "long-term host range" can evolve over time. Previously, the BHR multidrug resistance plasmid pB10 was shown to be highly unstable in Stenotrophomonas maltophilia P21 and Pseudomonas putida H2. To investigate whether this plasmid can adapt to such unfavorable hosts, we performed evolution experiments wherein pB10 was maintained in strain P21, strain H2, and alternatingly in P21 and H2. Plasmids that evolved in P21 and in both hosts showed increased stability and decreased cost in ancestral host P21. However, the latter group showed higher variability in stability patterns, suggesting that regular switching between distinct hosts hampered adaptive plasmid evolution. The plasmids evolved in P21 were also equally or more stable in other hosts compared to pB10, which suggested true host-range expansion. The complete genome sequences of four evolved plasmids with improved stability showed only one or two genetic changes. The stability of plasmids evolved in H2 improved only in their coevolved hosts, not in the ancestral host. Thus a BHR plasmid can adapt to an unfavorable host and thereby expand its long-term host range.
Assuntos
Evolução Molecular , Interações Hospedeiro-Patógeno/genética , Plasmídeos/genética , Pseudomonas putida/genética , Stenotrophomonas/genética , Instabilidade Genômica , Mutação/genética , FilogeniaRESUMO
Although the production and stabilization of metal nanoparticles (MNPs) is well understood, the behavior of these MNPs (possible aggregation or disaggregation) when they are intentionally or unintentionally exposed to different environments is a factor that continues to be underrated or overlooked. A case study is performed to analyze the stability of silver nanoparticles (AgNPs)-one of the most frequently used MNPs with excellent antibacterial properties-within two bacterial growth media: a minimally defined medium (IDL) and an undefined complex medium (LB). Moreover, the effect of aging, size and stabilization mechanisms is considered. Results clearly indicate a strong aggregation when AgNPs are dispersed in IDL. Regarding LB, the 100 nm electrosterically stabilized AgNPs remain stable while all others aggregate. Moreover, a serious aging effect is observed for the 10 nm electrostatically stabilized AgNPs when added to LB: after aggregation a restabilization effect occurs over time. Generally, this study demonstrates that the aging, medium composition (environment), size and stabilization mechanism-rarely acknowledged as important factors in nanotoxicity studies-have a profound impact on the AgNPs stabilization and should gain more attention in scientific research.
RESUMO
Horizontal plasmid transfer plays a key role in bacterial adaptation. In harsh environments, bacterial populations adapt by sampling genetic material from a horizontal gene pool through self-transmissible plasmids, and that allows persistence of these mobile genetic elements. In the absence of selection for plasmid-encoded traits it is not well understood if and how plasmids persist in bacterial communities. Here we present three models of the dynamics of plasmid persistence in the absence of selection. The models consider plasmid loss (segregation), plasmid cost, conjugative plasmid transfer, and observation error. Also, we present a stochastic model in which the relative fitness of the plasmid-free cells was modeled as a random variable affected by an environmental process using a hidden Markov model (HMM). Extensive simulations showed that the estimates from the proposed model are nearly unbiased. Likelihood-ratio tests showed that the dynamics of plasmid persistence are strongly dependent on the host type. Accounting for stochasticity was necessary to explain four of seven time-series data sets, thus confirming that plasmid persistence needs to be understood as a stochastic process. This work can be viewed as a conceptual starting point under which new plasmid persistence hypotheses can be tested.
Assuntos
Bactérias/genética , Modelos Genéticos , Plasmídeos/genética , Técnicas de Transferência de Genes , Cadeias de Markov , Seleção GenéticaRESUMO
Numerous studies have investigated the antibacterial effect of both silver ions and silver nanomaterials on a large diversity of environmentally and clinically relevant bacteria. However, contradictory results are reported in which inhibition concentrations were varying by a 10-fold. This study investigated whether this variance in results could be attributed to the difference in experimental conditions, especially the microbial growth medium. B. subtilis was exposed to 500 µg L-1 Ag+ in liquid growth media with different concentrations of some commonly used media components: tryptone, yeast extract, Cl-, and S2-. The toxic effect was investigated by means of three complementary analysis techniques: (i) analyzing the growth curves obtained by optical density measurements, (ii) using flow cytometry, and (iii) by transmission electron microscopy. The silver ion toxicity towards B. subtilis decreased as more tryptone, yeast extract, or S2- was present. This study demonstrates that the medium composition, rarely acknowledged as an important experimental factor in bacterial toxicity studies, has a profound impact on the observed silver toxicity towards B. subtilis.
Assuntos
Antibacterianos/química , Antibacterianos/farmacologia , Bacillus subtilis/efeitos dos fármacos , Nanopartículas Metálicas/química , Prata/química , Prata/farmacologia , Citometria de Fluxo , Concentração de Íons de Hidrogênio , Microscopia Eletrônica de TransmissãoRESUMO
Mycotoxins are toxic metabolites produced by fungi. To mitigate mycotoxins in food or feed, biotransformation is an emerging technology in which microorganisms degrade toxins into non-toxic metabolites. To monitor deoxynivalenol (DON) biotransformation, analytical tools such as ELISA and liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) are typically used. However, these techniques do not give a decisive answer about the remaining toxicity of possible biotransformation products. Hence, a bioassay using Lemna minor L. was developed. A dose-response analysis revealed significant inhibition in the growth of L. minor exposed to DON concentrations of 0.25 mg/L and higher. Concentrations above 1 mg/L were lethal for the plant. This bioassay is far more sensitive than previously described systems. The bioassay was implemented to screen microbial enrichment cultures, originating from rumen fluid, soil, digestate and activated sludge, on their biotransformation and detoxification capability of DON. The enrichment cultures originating from soil and activated sludge were capable of detoxifying and degrading 5 and 50 mg/L DON. In addition, the metabolites 3-epi-DON and the epimer of de-epoxy-DON (3-epi-DOM-1) were found as biotransformation products of both consortia. Our work provides a new valuable tool to screen microbial cultures for their detoxification capacity.
Assuntos
Araceae/metabolismo , Bioensaio/métodos , Microbiologia de Alimentos , Esgotos/microbiologia , Carneiro Doméstico/microbiologia , Microbiologia do Solo , Tricotecenos/metabolismo , Animais , Araceae/efeitos dos fármacos , Araceae/crescimento & desenvolvimento , Bioensaio/normas , Calibragem , Cromatografia Líquida , Relação Dose-Resposta a Droga , Ensaios de Triagem em Larga Escala , Inativação Metabólica , Cinética , Padrões de Referência , Espectrometria de Massas em Tandem , Tricotecenos/toxicidadeRESUMO
Exposure to mycotoxins, secondary metabolites produced by fungi, may infer serious risks for animal and human health and lead to economic losses. Several approaches to reduce these mycotoxins have been investigated such as chemical removal, physical binding, or microbial degradation. This review focuses on the microbial degradation or transformation of mycotoxins, with specific attention to the actual detoxification mechanisms of the mother compound. Furthermore, based on the similarities in chemical structure between groups of mycotoxins and environmentally recalcitrant compounds, known biodegradation pathways and degrading organisms which hold promise for the degradation of mycotoxins are presented.
RESUMO
Temporarily discontinuing the use of antibiotics has been proposed as a means to eliminate resistant bacteria by allowing sensitive clones to sweep through the population. In this study, we monitored a tetracycline-sensitive subpopulation that emerged during experimental evolution of E. coli K12 MG1655 carrying the multiresistance plasmid pB10 in the absence of antibiotics. The fraction of tetracycline-sensitive mutants increased slowly over 500 generations from 0.1 to 7%, and loss of resistance could be attributed to a recombination event that caused deletion of the tet operon. To help understand the population dynamics of these mutants, three mathematical models were developed that took into consideration recurrent mutations, increased host fitness (selection), or a combination of both mechanisms (full model). The data were best explained by the full model, which estimated a high mutation frequency (lambda = 3.11 x 10(-5)) and a significant but small selection coefficient (sigma = 0.007). This study emphasized the combined use of experimental data, mathematical models, and statistical methods to better understand and predict the dynamics of evolving bacterial populations, more specifically the possible consequences of discontinuing the use of antibiotics.