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1.
Int J Mol Sci ; 22(13)2021 Jun 24.
Artigo em Inglês | MEDLINE | ID: mdl-34202675

RESUMO

Garlic (Allium sativum) is a perennial bulbous plant. Due to its clonal propagation, various diseases threaten the yield and quality of garlic. In this study, we conducted in silico analysis to identify microorganisms, bacteria, fungi, and viruses in six different tissues using garlic RNA-sequencing data. The number of identified microbial species was the highest in inflorescences, followed by flowers and bulb cloves. With the Kraken2 tool, 57% of identified microbial reads were assigned to bacteria and 41% were assigned to viruses. Fungi only made up 1% of microbial reads. At the species level, Streptomyces lividans was the most dominant bacteria while Fusarium pseudograminearum was the most abundant fungi. Several allexiviruses were identified. Of them, the most abundant virus was garlic virus C followed by shallot virus X. We obtained a total of 14 viral genome sequences for four allexiviruses. As we expected, the microbial community varied depending on the tissue types, although there was a dominant microorganism in each tissue. In addition, we found that Kraken2 was a very powerful and efficient tool for the bacteria using RNA-sequencing data with some limitations for virome study.


Assuntos
Alho/microbiologia , Metagenoma , Metagenômica , Microbiota , Bactérias/classificação , Bactérias/genética , Biologia Computacional/métodos , Metagenômica/métodos , Especificidade de Órgãos , Filogenia , Análise de Sequência de RNA
2.
Int J Mol Sci ; 22(13)2021 Jun 22.
Artigo em Inglês | MEDLINE | ID: mdl-34206508

RESUMO

Plants of the genus Allium developed a diversity of defense mechanisms against pathogenic fungi of the genus Fusarium, including transcriptional activation of pathogenesis-related (PR) genes. However, the information on the regulation of PR factors in garlic (Allium sativum L.) is limited. In the present study, we identified AsPR genes putatively encoding PR1, PR2, PR4, and PR5 proteins in A. sativum cv. Ershuizao, which may be involved in the defense against Fusarium infection. The promoters of the AsPR1-5 genes contained jasmonic acid-, salicylic acid-, gibberellin-, abscisic acid-, auxin-, ethylene-, and stress-responsive elements associated with the response to plant parasites. The expression of AsPR1c, d, g, k, AsPR2b, AsPR5a, c (in roots), and AsPR4a(c), b, and AsPR2c (in stems and cloves) significantly differed between garlic cultivars resistant and susceptible to Fusarium rot, suggesting that it could define the PR protein-mediated protection against Fusarium infection in garlic. Our results provide insights into the role of PR factors in A. sativum and may be useful for breeding programs to increase the resistance of Allium crops to Fusarium infections.


Assuntos
Fusarium , Alho/genética , Alho/microbiologia , Genes de Plantas , Interações Hospedeiro-Patógeno/genética , Família Multigênica , Doenças das Plantas/genética , Doenças das Plantas/microbiologia , Clonagem Molecular , Fusarium/fisiologia , Regulação da Expressão Gênica de Plantas , Genoma de Planta , Genômica/métodos , Regiões Promotoras Genéticas
3.
PLoS One ; 16(5): e0250571, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33989289

RESUMO

Soil microbial communities are affected by interactions between agricultural management (e.g., fertilizer) and soil compartment, but few studies have considered combinations of these factors. We compared the microbial abundance, diversity and community structure in two fertilizer dose (high vs. low NPK) and soil compartment (rhizosphere vs. bulk soils) under 6-year fertilization regimes in a continuous garlic cropping system in China. The soil contents of NO3- and available K were significantly higher in bulk soil in the high-NPK. The 16S rRNA gene-based bacterial and archaeal abundances were positively affected by both the fertilizer dose and soil compartment, and were higher in the high-NPK fertilization and rhizosphere samples. High-NPK fertilization increased the Shannon index and decreased bacterial and archaeal richness, whereas the evenness was marginally positively affected by both the fertilizer dose and soil compartment. Soil compartment exerted a greater effect on the bacterial and archaeal community structure than did the fertilization dose, as demonstrated by both the nonmetric multidimensional scaling and redundancy analysis results. We found that rhizosphere effects significantly distinguished 12 dominant classes of bacterial and archaeal communities, whereas the fertilizer dose significantly identified four dominant classes. In particular, a Linear Effect Size analysis showed that some taxa, including Alphaproteobacteria, Rhizobiales, Xanthomonadaceae and Flavobacterium, were enriched in the garlic rhizosphere of the high-NPK fertilizer samples. Overall, the fertilizer dose interacted with soil compartment to shape the bacterial and archaeal community composition, abundance, and biodiversity in the garlic rhizosphere. These results provide an important basis for further understanding adaptive garlic-microbe feedback, reframing roots as a significant moderating influence in agricultural management and shaping the microbial community.


Assuntos
Archaea/genética , Bactérias/genética , Fertilizantes/análise , Alho/crescimento & desenvolvimento , Rizosfera , Microbiologia do Solo/normas , Solo/química , Archaea/isolamento & purificação , Bactérias/isolamento & purificação , Biodiversidade , China , Alho/genética , Alho/microbiologia , RNA Ribossômico 16S/genética
4.
Compr Rev Food Sci Food Saf ; 19(2): 801-834, 2020 03.
Artigo em Inglês | MEDLINE | ID: mdl-33325167

RESUMO

Black garlic (BG) is essentially a processed food and obtained through the transformation of fresh garlic (FG) (Allium sativum L.) via a range of chemical reactions (including the Maillard reaction) and microbial fermentation. This review provides the up-to-date knowledge of the dynamic and complicated changes in major components during the conversion of FG to BG, including moisture, lipids, carbohydrates (such as sugars), proteins, organic acids, organic sulfur compounds, alkaloids, polyphenols, melanoidins, 5-hydroxymethylfurfural, vitamins, minerals, enzymes, and garlic endophytes. The obtained evidence confirms that BG has several advantages over FG in certain product attributes and biological properties (especially antioxidant activity), and the factors affecting the quality of BG include the type and characteristics of FG and processing technologies and methods (especially pretreatments, and processing temperature and humidity). The interactions among garlic components, and between garlic nutrients and microbes, as well as the interplay between pretreatment and main manufacturing process, all determine the sensory and nutritional qualities of BG. Before BG is marketed as a novel snack or functional food, more research is required to fill the knowledge gaps related to quantitative monitoring of the changes in metabolites (especially those taste-active and/or biological-active substances) during BG manufacturing to maximize BG's antioxidant, anticancer, antiobesity, anti-inflammatory, immunostimulatory, anti-allergic, hepatoprotective, cardioprotective and oxidative stress-/hangover syndrome-reducing functions, and beneficial effects on memory/nervous systems. Assessments of the quality, efficacy, and safety of BG should be performed considering the impacts of BG production conditions, postproduction handling, and intake methods.


Assuntos
Alho/química , Valor Nutritivo , Antioxidantes/química , Fermentação , Manipulação de Alimentos/métodos , Alho/microbiologia , Reação de Maillard
5.
Food Funct ; 11(12): 10913-10924, 2020 Dec 01.
Artigo em Inglês | MEDLINE | ID: mdl-33242049

RESUMO

Fermenting garlic and onion provides the advantages of storage life extension, anti-oxidative and anti-diabetic activities, and their metabolite, allyl mercaptan, offers a strong aroma and various health benefits. Here, we report the probiotic properties of Lactobacillus pentosus SMB718 isolated from Korean traditional paste and its high allyl mercaptan productivity in garlic and onion fermentation. This strain was safe for use in food fermentation, as it was a non-biogenic amine producer and non-hemolytic. It showed high stability under simulated human gastrointestinal conditions and good adhesion ability to intestinal epithelial cells, including both Caco-2 and HT-29. This strain had antioxidant and anti-microbial activities. In addition, the heat-killed cells and lysate exerted anti-inflammatory effects on both LPS-stimulated RAW 264.7 cells and mouse macrophages by inhibition of pro-inflammatory cytokines and induction of anti-inflammatory cytokines. Furthermore, this strain possessed good fermentation properties in garlic and onion-enriched radish juice (GORJ); it grew well decreasing the pH and provided a rich aroma compound during fermentation. When L. pentosus SMB718 was used as a starter in garlic and onion fermentation, a significantly higher amount of allyl mercaptan (344 ppb) was produced compared with that produced by the type strain (82 ppb). In conclusion, L. pentosus SMB718 can be used as a beneficial probiotic starter for better flavor production in the fermentation of allium species plants.


Assuntos
Fermentação , Alimentos Fermentados , Alho/química , Lactobacillus pentosus/fisiologia , Cebolas/química , Probióticos , Compostos de Sulfidrila/metabolismo , Animais , Anti-Infecciosos/farmacologia , Anti-Inflamatórios , Antioxidantes/farmacologia , Células CACO-2 , Citocinas , Células Epiteliais , Alho/microbiologia , Células HT29 , Humanos , Camundongos , Cebolas/microbiologia , Células RAW 264.7
6.
Microbiol Res ; 241: 126585, 2020 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-32919224

RESUMO

Garlic (Allium sativum L.) is one of the crops whose economic importance has increased considerably in recent years in Brazil. The use of plant growth-promoting bacteria (PGPB) is a useful alternative for reducing the cost of agricultural inputs and, consequently, for increasing productivity. Therefore, the main objective of this work was to isolate and evaluate potential growth promoters in plants and plant pathogenic fungi growth inhibitors using endophytic bacteria from garlic roots and bacteria from the Agricultural Microbiology Culture Collection at the Federal University of Lavras. Besides verifying improvements in the growth and physiology of garlic meristems grown in vitro under the action of PGPB. Forty-eight endophytic bacteria were identified using matrix-assisted laser desorption/ionization-time of flight mass spectrometry based on the protein profile of each isolate. Four isolates were chosen according to their ability to fix nitrogen, to produce auxin and solubilize phosphate. The cultivation of garlic meristems in tissue culture with these bacteria was established at a population level of 106 CFU/mL. The evaluated criteria were: (1) the colonization capacity of the bacteria inside the garlic plants determined through scanning electron microscopy; (2) the chlorophyll content; and (3) the growth of garlic plants in vitro post-PGPB inoculation. Volatiles emitted by those isolates inhibited fungi growth. The inoculation of garlic meristems with Enterobacter cloacae and Burkholderia cepacia promoted an improvement in the growth and physiological attributes of garlic, indicating the feasibility of their application as plant growth promoters for commercial cultivation.


Assuntos
Bactérias/metabolismo , Alho/microbiologia , Meristema/crescimento & desenvolvimento , Desenvolvimento Vegetal/fisiologia , Raízes de Plantas/crescimento & desenvolvimento , Antifúngicos/metabolismo , Bactérias/isolamento & purificação , Brasil , Burkholderia cepacia/metabolismo , Clorofila/análise , Endófitos/metabolismo , Enterobacter cloacae/metabolismo , Ácidos Indolacéticos/metabolismo , Microscopia Eletrônica de Varredura , Fixação de Nitrogênio/fisiologia , Raízes de Plantas/microbiologia
7.
Food Chem ; 318: 126481, 2020 Jul 15.
Artigo em Inglês | MEDLINE | ID: mdl-32126467

RESUMO

Fermented foods constitute hubs of microbial consortia differentially affecting nutritional and organoleptic properties, quality, and safety. Here we show the origin source of fermentative microbes and fermentation dynamics of kimchi. We partitioned microbiota by raw ingredient (kimchi cabbage, garlic, ginger, and red pepper) to render kimchi fermented by each source-originated microbe pool and applied multi-omics (metataxonomics and metabolomics), bacterial viability, and physiochemical analyses to longitudinally collected samples. Only kimchi cabbage- and garlic-derived microbial inoculums yielded successful kimchi fermentations. The dominant fermentative microbial taxa and subsequent metabolic outputs differed by raw ingredient type: the genus Leuconostoc, Weissella, and Lactobacillus for all non-sterilized ingredients, garlic, and kimchi cabbage, respectively. Gnotobiotic kimchi inoculated by mono-, di-, and tri- isolated fermentative microbe combinations further revealed W. koreensis-mediated reversible microbial metabolic outputs. The results suggest that the raw ingredient microbial habitat niches selectively affect microbial community assembly patterns and processes during kimchi fermentation.


Assuntos
Alimentos Fermentados/microbiologia , Microbiota , Brassica/microbiologia , Capsicum/microbiologia , Fermentação , Microbiologia de Alimentos , Alho/microbiologia , Zingiber officinale/microbiologia , Lactobacillus/genética , Leuconostoc/genética , Metaboloma , Consórcios Microbianos , Microbiota/genética , Weissella/genética
8.
PLoS One ; 15(3): e0228485, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32130211

RESUMO

Fifty four Trichoderma strains were isolated from soil samples collected from garlic and onion crops in eight different sites in Brazil and were identified using phylogenetic analysis based on combined ITS region, tef1-α, cal, act and rpb2 sequences. The genetic variability of the recovered Trichoderma species was analysed by AFLP and their phenotypic variability determined using MALDI-TOF. The strain clusters from both typing techniques coincided with the taxonomic determinations made from phylogenetic analysis. The phylogenetic analysis showed the occurrence of Trichoderma asperellum, Trichoderma asperelloides, Trichoderma afroharzianum, Trichoderma hamatum, Trichoderma lentiforme, Trichoderma koningiopsis, Trichoderma longibrachiatum and Trichoderma erinaceum, in the soil samples. We also identified and describe two new Trichoderma species, both in the harzianum clade of section Pachybasium, which we have named Trichoderma azevedoi sp. nov. and Trichoderma peberdyi sp. nov. The examined strains of both T. azevedoi (three strains) and T. peberdyi (12 strains) display significant genotypic and phenotypic variability, but form monophyletic clades with strong bootstrap and posterior probability support and are morphologically distinct from their respective most closely related species.


Assuntos
Alho/microbiologia , Cebolas/microbiologia , Microbiologia do Solo , Trichoderma/classificação , Trichoderma/isolamento & purificação , Análise do Polimorfismo de Comprimento de Fragmentos Amplificados , Biodiversidade , Brasil , DNA Fúngico/análise , DNA Fúngico/genética , Técnicas de Tipagem Micológica/métodos , Filogenia , Análise de Sequência de DNA/métodos , Especificidade da Espécie , Trichoderma/citologia , Trichoderma/genética
9.
Curr Microbiol ; 77(7): 1150-1158, 2020 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-32060767

RESUMO

The garlic contains sulfur bioactive compounds responsible for medicinal properties. The decrease of these compounds due to inadequate storage conditions reduces the beneficial properties and favors infection by microorganisms. Several studies have shown high frequency of garlic infected with Aspergillus section Nigri that potentially produce mycotoxin. Garlic samples were collected in markets of Brazil and a total of 32 samples (of 36) had the fungal infection with predominant genus Aspergillus (50.3%), Penicillium (34.7%), and Fusarium (11%). A total of 63% (649/1031) of infection with Aspergillus section Nigri, of which 60 isolates were selected for analysis of genetic variability that resulted in 4 clusters. Representatives of clusters were identified by the calmodulin gene. Isolates from cluster I were subdivided into A-I and identified as A. niger (16 isolates) and the isolates of clusters B-I, II, and III were identified as A. welwitschiae (43 isolates). Besides, an isolate of the IV-cluster was identified by A. luchuensis. Further, we used the multiplex PCR to verify genotypes of 59 isolates, and none of these had OTA production-associated genotype. Moreover, 19 A. welwitschiae and 15 A. niger were FB2 production-associated genotype. Our study is the first report to the incidence of garlic infection in Brazil and to show that A. welwitschiae causes most of these infections.


Assuntos
Aspergillus , Fumonisinas/metabolismo , Alho/microbiologia , Ocratoxinas/metabolismo , Aspergillus/genética , Aspergillus/metabolismo , Aspergillus/patogenicidade , Brasil , Microbiologia de Alimentos , Genótipo
10.
Int J Food Microbiol ; 316: 108440, 2020 Mar 02.
Artigo em Inglês | MEDLINE | ID: mdl-31733520

RESUMO

Salmonella is the causative agent of many outbreaks related to spice consumption. However, because of the antimicrobial properties of various spices which hinders recovery and detection, Salmonella detection in spices remains a challenge. The objective of this study was to optimize an enrichment broth for Salmonella growth in different spices and tea, in order to maintain an adequate pH and decrease the antimicrobial effects of spices during Salmonella enrichment and subsequent detection. Salmonella contaminated spice and tea dried samples were prepared and the detection of Salmonella was assessed using the developed broth and automated DNA extraction and RT-PCR. Double strength Buffered Peptone Water (BPW) was used to maintain pH, and L-cysteine and DL-serine were added to the broth to reduce the effects of antimicrobial compounds in spices. The modified enrichment broth allowed the growth of Salmonella from each spice sample. Sample to broth ratios varied from 1:9 (garlic powder, chili peppers and tea), to 1:20 (cinnamon). The pH value of each enrichment varied but remained above 4.8. The addition of L-cysteine (30 mmol/L) allowed Salmonella recovery and growth in garlic and onion samples and the addition of DL-serine (11.23 mmol/L) allowed the recovery and growth in cinnamon. The results indicated that Salmonella detection was achieved in <24 h in the modified (BPW + L-cysteine and DL-serine) enrichment broth followed by detection by RT-PCR. This protocol could allow for a more rapid, robust, and sensitive enrichment method for Salmonella in spices.


Assuntos
Microbiologia de Alimentos/métodos , Salmonella/isolamento & purificação , Especiarias/microbiologia , Chá/microbiologia , Capsicum/microbiologia , Cinnamomum zeylanicum/microbiologia , Meios de Cultura/química , Alimentos em Conserva/microbiologia , Alho/microbiologia , Cebolas/microbiologia , Salmonella/genética , Salmonella/crescimento & desenvolvimento
11.
Hig. aliment ; 33(288/289): 2343-2347, abr.-maio 2019. tab
Artigo em Português | LILACS, VETINDEX | ID: biblio-1482216

RESUMO

O risco de Doenças Veiculadas por Alimentos - DVA em hambúrgueres pode ser minimizado pelo uso de antimicrobianos naturais. O objetivo do presente trabalho foi avaliar a eficiência de diferentes concentrações do Extrato Aquoso de Alho - EAA na inibição de Escherichia coli, Salmonella enterica, Cronobacter sakazakii, Listeria monocytogenes, Staphylococcus aureus e Bacillus cereus e avalia r sensorialmente as amostras de hambúrguer adicionadas de EAA. Os resultados indicaram que o EAA inibiu o crescimento das cepas bacterianas, sendo a ação inibitória proporcional às concentrações usadas sendo que a refrigeração a 7°C por 72 h do EAA não afetou seu efeito inibitório. Na análise sensorial, houve boa aceitabilidade e intenção de compra positiva para as amostras, com destaque para aquelas com maior concentração de EAA (1000μL/mL).


Assuntos
Alho/microbiologia , Antibacterianos/análise , Comportamento do Consumidor/estatística & dados numéricos , Produtos da Carne/análise , Produtos da Carne/microbiologia , Técnicas Bacteriológicas/análise , Doenças Transmitidas por Alimentos
12.
An Acad Bras Cienc ; 90(4): 3949-3962, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30427392

RESUMO

Garlic (Allium sativum L.) plays an important role in popular culture due to its dietary and medicinal uses. It is also used to produce a wide range of pharmacologically interesting molecules. Several pathogens affect garlic plants, especially Athelia (Sclerotium) rolfsii, a fungus that is widespread and causes large economic losses. It causes direct damage to crops and leads to plant stress, which induces secondary metabolite production in plants. The use of microorganisms as biocontrol agents may induce the production of beneficial metabolites in plants that will protect it and promote resistance to pathogen attack. In addition to suppressing disease, biological control agents may have elicitor effects that could induce an increase in the production of useful bioactive secondary metabolites in plants, some of which may be of pharmacological interest. Therefore, the search for new biological control agents should also consider their potential as elicitor agents. This paper presents an analysis of the biological control of Athelia (Sclerotium) rolfsii by antagonistic microrganisms, the potential of yeasts and bacteria of the genus Bacillus for the biocontrol of phytopathogens, microrganisms influence in nutritional and bioactive compounds content of interest to the pharmaceutical industry.


Assuntos
Basidiomycota/patogenicidade , Agentes de Controle Biológico , Alho/microbiologia , Doenças das Plantas/microbiologia , Doenças das Plantas/prevenção & controle
13.
Sci Rep ; 8(1): 14514, 2018 09 28.
Artigo em Inglês | MEDLINE | ID: mdl-30266995

RESUMO

Antimicrobial genes are found in all classes of life. To efficiently isolate these genes, we used Bacillus subtilis and Escherichia coli as target indicator bacteria and transformed them with cDNA libraries. Among thousands of expressed proteins, candidate proteins played antimicrobial roles from the inside of the indicator bacteria (internal effect), contributing to the sensitivity (much more sensitivity than the external effect from antimicrobial proteins working from outside of the cells) and the high throughput ability of screening. We found that B. subtilis is more efficient and reliable than E. coli. Using the B. subtilis expression system, we identified 19 novel, broad-spectrum antimicrobial genes. Proteins expressed by these genes were extracted and tested, exhibiting strong external antibacterial, antifungal and nematicidal activities. Furthermore, these newly isolated proteins could control plant diseases. Application of these proteins secreted by engineered B. subtilis in soil could inhibit the growth of pathogenic bacteria. These proteins are thermally stable and suitable for clinical medicine, as they exhibited no haemolytic activity. Based on our findings, we speculated that plant, animal and human pathogenic bacteria, fungi or even cancer cells might be taken as the indicator target cells for screening specific resistance genes.


Assuntos
Bacillus subtilis/genética , Resistência à Doença/genética , Alho/genética , Pinellia/genética , Proteínas de Plantas/genética , Animais , Bacillus subtilis/metabolismo , Caenorhabditis elegans , Membrana Celular/ultraestrutura , Clonagem Molecular , DNA Complementar/genética , DNA de Plantas/genética , Escherichia coli/genética , Escherichia coli/metabolismo , Alho/microbiologia , Bactérias Gram-Negativas , Bactérias Gram-Positivas , Ensaios de Triagem em Larga Escala , Interações Hospedeiro-Patógeno , Organismos Geneticamente Modificados , Pinellia/microbiologia , Doenças das Plantas/microbiologia , Doenças das Plantas/prevenção & controle , Proteínas de Plantas/isolamento & purificação , Proteínas de Plantas/fisiologia , Estabilidade Proteica , Proteínas Recombinantes/genética , Especificidade da Espécie , Transformação Bacteriana
14.
Food Res Int ; 106: 428-438, 2018 04.
Artigo em Inglês | MEDLINE | ID: mdl-29579944

RESUMO

Black garlic is a distinctive garlic deep-processed product made from fresh garlic at high temperature and controlled humidity. To explore microbial community structure, diversity and metabolic potential during the 12days of the black garlic processing, Illumina MiSeq sequencing technology was performed to sequence the 16S rRNA V3-V4 hypervariable region of bacteria. A total of 677,917 high quality reads were yielded with an average read length of 416bp. Operational taxonomic units (OTU) clustering analysis showed that the number of species OTUs ranged from 148 to 1974, with alpha diversity increasing remarkably, indicating the high microbial community abundance and diversity. Taxonomic analysis indicated that bacterial community was classified into 45 phyla and 1125 distinct genera, and the microbiome of black garlic samples based on phylogenetic analysis was dominated by distinct populations of four genera: Thermus, Corynebacterium, Streptococcus and Brevundimonas. The metabolic pathways were predicted for 16S rRNA marker gene sequences based on Kyoto Encyclopedia of Genes and Genomes (KEGG), indicating that amino acid metabolism, carbohydrate metabolism and membrane transport were important for the black garlic fermentation process. Overall, the study was the first to reveal microbial community structure and speculate the composition of functional genes in black garlic samples. The results contributed to further analysis of the interaction between microbial community and black garlic components at different stages, which was of great significance to study the formation mechanism and quality improvement of black garlic in the future.


Assuntos
Bactérias/isolamento & purificação , Fermentação , Manipulação de Alimentos/métodos , Microbiologia de Alimentos/métodos , Alho/microbiologia , Microbiota , Raízes de Plantas/microbiologia , Bactérias/classificação , Bactérias/genética , Bactérias/metabolismo , Corynebacterium/genética , Corynebacterium/isolamento & purificação , Corynebacterium/metabolismo , Fermentação/genética , Sequenciamento de Nucleotídeos em Larga Escala , Temperatura Alta , Umidade , Microbiota/genética , Filogenia , Ribotipagem , Streptococcus/genética , Streptococcus/isolamento & purificação , Streptococcus/metabolismo , Thermus/genética , Thermus/isolamento & purificação , Thermus/metabolismo
15.
Microbiologyopen ; 7(1)2018 02.
Artigo em Inglês | MEDLINE | ID: mdl-28990361

RESUMO

The objectives of this study were to isolate and identify garlic endophytes, and explore the characteristics of dominant strains. Garlic endophytes were studied through phenotypical characterization and comparative sequence analysis of 16S rDNA based on culture-dependent approaches. Representative strains inferred from 16S rDNA sequencing were selected for further identification by gyrA and rpoB gene loci and phylogenetic analysis based on concateneted house-keeping sequences. Seven kinds of Bacillus were found from garlic and black garlic, respectively. Further studies demonstrated that the total bacteria and endophytes showed a sharp decrease firstly, followed by a rapid rise, then maintained at a certain level, and finally slowed down during the black garlic processing. B. subtilis, B. methylotrophicus, and B. amyloliquefaciens were the dominant strains. The selected strains were capable of fermenting glucose, lactose, sucrose, and garlic polysaccharide to produce acid but no gas, with a strong ability of heat resistance. The results indicated that there were a certain number of garlic endophytes during the black garlic processing, and Bacillus was the dominant strains under the conventional culture-dependent methods. This report provided useful information for the presence and type of garlic endophytes during the black garlic processing, which were of great significance to study the formation mechanism and quality improvement of black garlic in the future, as well as the security of garlic powder.


Assuntos
Bacillus/classificação , Bacillus/isolamento & purificação , Endófitos/classificação , Endófitos/isolamento & purificação , Alho/microbiologia , Bacillus/genética , Bacillus/fisiologia , Técnicas de Tipagem Bacteriana , Análise por Conglomerados , DNA Girase/genética , DNA Ribossômico/química , DNA Ribossômico/genética , RNA Polimerases Dirigidas por DNA/genética , Endófitos/genética , Endófitos/fisiologia , Genes Essenciais , Filogenia , RNA Ribossômico 16S/genética , Análise de Sequência de DNA
16.
Int J Food Microbiol ; 263: 26-31, 2017 Dec 18.
Artigo em Inglês | MEDLINE | ID: mdl-29024904

RESUMO

Salmonella survival was characterized and modeled during thermal dehydration of fresh garlic and storage of dehydrated garlic products. In our experiments that simulated commercial dehydration processing at 80±5°C, moderate level of Salmonella contamination (4-5logCFU/g) on fresh garlic was reduced below the enumeration limit (1.7logCFU/g) after 4.5h of dehydration and not detectable by culture enrichment after 7h. With high level of contamination (7-8logCFU/g), the Salmonella population persisted at 3.6logCFU/g after 8h of processing. By increasing the dehydration temperature to 90±5°C, the moderate and high levels of initial Salmonella load on fresh garlic dropped below the enumeration limit after 1.5 and 3.75h of processing and became undetectable by culture enrichment after 2.5 and 6h, respectively. During the storage of dried garlic products, Salmonella was not able to grow under all tested combinations of temperature (25 and 35°C) and water activity (0.56-0.98) levels, suggesting active inhibition. Storage temperature played a primary role in determining Salmonella survival on dehydrated garlic flakes. Under a typical storage condition at 25°C and ambient relative humidity, Salmonella could persist over months with the population gradually declining (4.3 log reduction over 88days). Granular size of dehydrated garlic had an impact on Salmonella survival, with better survival of the pathogen observed in bigger granules. At the early stage of dehydrated garlic storage (until 7days), rising water activity appeared to initially promote but then inhibited Salmonella survival, resulting in a water activity threshold at 0.73 where Salmonella displayed strongest persistence. However, this phenomenon was less apparent during extended storage (after 14days).


Assuntos
Alho/microbiologia , Salmonella/crescimento & desenvolvimento , Contagem de Colônia Microbiana , Desidratação , Conservação de Alimentos , Armazenamento de Alimentos , Alho/química , Salmonella/química , Salmonella/metabolismo , Temperatura , Fatores de Tempo , Água/análise , Água/metabolismo
17.
Rev. iberoam. micol ; 34(3): 158-164, jul.-sept. 2017. tab
Artigo em Inglês | IBECS | ID: ibc-165194

RESUMO

Background. The colonization of the surfaces of dental prostheses by Candida albicans is associated with the development of denture stomatitis. In this context, the use of fluconazole has been proposed, but its disadvantage is microbial resistance. Meanwhile, the oil of Allium sativum has shown an effect in controlling biofilm formation by C. albicans. Aims. The objective of this study was to determine the antifungal activities of the essential oil of A. sativum and fluconazole against clinical isolates of Candida species obtained from rigid, acrylic-based partial or total dentures and to compare these agents’ effects on both biofilm and planktonic cells. Methods. A total of 48 clinical isolates obtained from the acrylic surface of partial or complete dentures were examined, and the following species were identified: C. albicans, Candida glabrata, Candida tropicalis, and Candida krusei. For each isolate, the antifungal activities of the essential oil of A. sativum and fluconazole against both biofilm and planktonic cells were evaluated using the Clinical & Laboratory Standards Institute (CLSI) M27-A3 method. The isolates were also evaluated by semiquantitative XTT reduction. Results. All planktonic Candida isolates were susceptible to the essential oil of A. sativum, whereas 4.2% were resistant to fluconazole. Regarding susceptibilities in biofilms, 43.8% of biofilms were resistant to A. sativum oil, and 91.7% were resistant to fluconazole. Conclusions. All planktonic cells of the different Candida species tested are susceptible to <1mg/ml A. sativum oil, and the majority are susceptible to fluconazole. Susceptibility decreases in biofilm cells, with increased resistance to fluconazole compared with A. sativum oil. The essential oil of A. sativum is thus active against clinical isolates of Candida species obtained from dentures, with effects on both biofilm and planktonic cells in vitro (AU)


Antecedentes. La colonización por parte de Candida albicans de las superficies de las prótesis dentales se asocia con el desarrollo de estomatitis. Se ha propuesto el uso de fluconazol, pero su desventaja es la resistencia microbiana. El aceite de Allium sativum ha mostrado su efectividad al controlar la formación de biopelícula de objetivos. Objetivos. Determinar la sensibilidad de cepas clínicas de especies de Candida, obtenidas de prótesis dentales parciales o totales rígidas de base acrílica, al aceite esencial de A. sativum y comparar su efecto en células planctónicas y en biopelícula. Métodos. Se incluyeron 48 cepas clínicas de la superficie acrílica de prótesis dentales totales o parciales, identificadas entre las siguientes especies: C. albicans, Candida glabrata, Candida tropicalis y Candida krusei. Se evaluó la sensibilidad de cada una al aceite esencial de A. sativum y al fluconazol mediante la metodología M27-A3 del CLSI, tanto sobre células planctónicas como en biopelícula, y mediante el método semicuantitativo de la reducción de XTT en el último caso. Resultados. Todas las cepas planctónicas de Candida fueron sensibles al aceite esencial de A.sativum, mientras que el 4,2% fue resistente al fluconazol. En cuanto a su sensibilidad en biopelícula, el 43,8% fue resistente a A. sativum y el 91,7% lo fue al fluconazol. Conclusiones. Todas las cepas en forma planctónica de las diferentes especies de Candida fueron sensibles a concentraciones inferiores a 1mg/ml del aceite esencial de A. sativum y en menor proporción a fluconazol. La sensibilidad disminuyó en las células en biopelícula, con mayor resistencia al fluconazol en comparación con el aceite esencial de A. sativum. Por tanto, el aceite esencial de A. sativum es activo frente a cepas clínicas de diferentes especies de Candida, obtenidas de dentaduras, con efectos en biopelícula y células planctónicas in vitro (AU)


Assuntos
Humanos , 25939/administração & dosagem , 25939/uso terapêutico , Alho/microbiologia , Prótese Dentária/microbiologia , Candida/isolamento & purificação , Candida albicans/isolamento & purificação , Candida glabrata/isolamento & purificação , Candida tropicalis/isolamento & purificação , Testes de Sensibilidade Microbiana/métodos , Sensibilidade e Especificidade , Fluconazol/uso terapêutico
18.
J Agric Food Chem ; 65(32): 7000-7005, 2017 Aug 16.
Artigo em Inglês | MEDLINE | ID: mdl-28719747

RESUMO

An epidemic fungal disease caused by Fusarium proliferatum, responsible for fumonisin production (FB1, FB2, and FB3), has been reported in the main garlic-producing countries in recent years. Fumonisins are a group of structurally related toxic metabolites produced by this pathogen. The aim of this work was to establish an enzyme-linked immunosorbent assay (ELISA) procedure, mostly applied to cereals, that is suitable for fumonisin detection in garlic and compare these results to those obtained by high-performance liquid chromatography (HPLC) and screening of fresh and dehydrated garlic for toxicological risk. The results show good correlation between the two analytical methods. In fresh symptomatic garlic, fumonisin levels were higher in the basal plates than those in the portions with necrotic spots. Among the 56 commercially dehydrated garlic samples screened, three were positive by ELISA test and only one was above the limit of quantitation. The same samples analyzed by HPLC showed the presence of FB1 in trace amounts that was below the limit of quantitation; FB2 and FB3 were absent. The results are reassuring, because no substantial contamination by fumonisins was found in commercial garlic.


Assuntos
Contaminação de Alimentos/análise , Fumonisinas/análise , Alho/química , Micotoxinas/análise , Contaminação de Alimentos/economia , Manipulação de Alimentos , Fumonisinas/metabolismo , Fusarium/metabolismo , Alho/microbiologia , Micotoxinas/metabolismo , Doenças das Plantas/microbiologia
19.
J Food Sci ; 82(7): 1631-1639, 2017 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-28613443

RESUMO

Development and selection of model foods is a critical part of microwave thermal process development, simulation validation, and optimization. Previously developed model foods for pasteurization process evaluation utilized Maillard reaction products as the time-temperature integrators, which resulted in similar temperature sensitivity among the models. The aim of this research was to develop additional model foods based on different time-temperature integrators, determine their dielectric properties and color change kinetics, and validate the optimal model food in hot water and microwave-assisted pasteurization processes. Color, quantified using a* value, was selected as the time-temperature indicator for green pea and garlic puree model foods. Results showed 915 MHz microwaves had a greater penetration depth into the green pea model food than the garlic. a* value reaction rates for the green pea model were approximately 4 times slower than in the garlic model food; slower reaction rates were preferred for the application of model food in this study, that is quality evaluation for a target process of 90 °C for 10 min at the cold spot. Pasteurization validation used the green pea model food and results showed that there were quantifiable differences between the color of the unheated control, hot water pasteurization, and microwave-assisted thermal pasteurization system. Both model foods developed in this research could be utilized for quality assessment and optimization of various thermal pasteurization processes.


Assuntos
Manipulação de Alimentos , Qualidade dos Alimentos , Alho/química , Pasteurização , Pisum sativum/química , Clostridium botulinum/isolamento & purificação , Cor , Contaminação de Alimentos/análise , Microbiologia de Alimentos , Alho/microbiologia , Temperatura Alta , Micro-Ondas , Modelos Teóricos , Pisum sativum/microbiologia , Reprodutibilidade dos Testes , Temperatura
20.
Food Microbiol ; 67: 41-48, 2017 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-28648292

RESUMO

Fusarium proliferatum is a world-wide occurring fungal pathogen affecting several crops included garlic bulbs. In Spain, this is the most frequent pathogenic fungus associated with garlic rot during storage. Moreover, F. proliferatum is an important mycotoxigenic species, producing a broad range of toxins, which may pose a risk for food safety. The aim of this study is to assess the intraspecific variability of the garlic pathogen in Spain implied by analyses of translation elongation factor (tef-1α) and FUM1 gene sequences as well as the differences in growth rates. Phylogenetic characterization has been complemented with the characterization of mating type alleles as well as the species potential as a toxin producer. Phylogenetic trees based on the sequence of the translation elongation factor and FUM1 genes from seventy nine isolates from garlic revealed a considerable intraspecific variability as well as high level of diversity in growth speed. Based on the MAT alleles amplified by PCR, F. proliferatum isolates were separated into different groups on both trees. All isolates collected from garlic in Spain proved to be fumonisin B1, B2, and B3 producers. Quantitative analyses of fumonisins, beauvericin and moniliformin (common secondary metabolites of F. proliferatum) showed no correlation with phylogenetic analysis neither mycelial growth. This pathogen presents a high intraspecific variability within the same geographical region and host, which is necessary to be considered in the management of the disease.


Assuntos
Fusarium/genética , Fusarium/isolamento & purificação , Alho/microbiologia , Micotoxinas/metabolismo , Doenças das Plantas/microbiologia , Proteínas Fúngicas/genética , Proteínas Fúngicas/metabolismo , Fusarium/metabolismo , Variação Genética , Filogenia , Espanha
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