Choice of DNA extraction protocols from Gram negative and positive bacteria and directly from the soil

Staphylococcus aureus is an important pathogen involved in subclinical bovine mastitis, causing high economic losses for the dairy industry. The successful persistence of this pathogen in the host occurs due to a series of factors associated with colonization ability and the acquisition of virulence factors. This bacterial species carries genetic heterogeneity, and genetically diverse strains characterize the population. Analysis of the genetic variation is an important tool for epidemiological studies. For this study, S. aureus strains were randomly selected by molecular profiling. All strains were originated from the milk of cows of subclinical mastitis on farms in the State of Rio de Janeiro. Strains of S. aureus were profiled using virulence gene analysis profiles, agr and spa typing, Pulsed Field Gel Electrophoresis (PFGE), and Multilocus Sequence Typing (MLST). 47% (8/17) of the tested strains were positive strains for icaA gene; 82.3% (14/17) for icaD gene; 41% (7/17) for fbnA gene; 47% (8/17) positive for fbnB gene; 94% (16/17) for the hlA gene and 70.5% (12/17) for hlB gene. These virulence results generated 11 different profiles. Most strains (58.8% -10/17) were classified as type-II by agr system. spa typing identified seven different spa types. PFGE analysis found extensive genetic heterogeneity and no clones were observed. MLST analysis generated five different types of ST/CC. Considering the results observed at the present study, a high genetic variety of S. aureus strains associated of the presence of different virulence factors justified the absence of clonal strains at the properties evaluated. Besides, that, the definition of S. aureus clonal strain as well dynamic population in not fully understood since there are a limited number of studies in S. aureus associated with bovine mastitis.

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“…Bacterial total DNA extraction was performed according to the protocol established by Tito et al (2015).…”

Section: Dna Extractionmentioning

confidence: 99%

Molecular characterization and genetic diversity of Staphylococcus aureus isolates of dairy production farms in Rio de Janeiro, Brazil

et al. 2021

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“…Para a extração do DNA bacteriano, foi realizada metodologia padronizada e estabelecida pelo Laboratório de Bacteriologia Veterinária da UFRRJ (TITO et al, 2015).…”

Section: Extração Do Dna Bacterianounclassified

The persistence of biofilm-associated antibiotic resistance of Staphylococcus aureus isolated from clinical bovine mastitis cases in Australia

et al. 2013

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The aim of this investigation was to determine the persistence of biofilm-associated antibiotic resistance developed by methicillin-sensitive Staphylococcus aureus (MSSA), of different capsular types, during biofilm formation. Because of superiority of the tissue culture plate (TCP) over the Congo Red Agar (CRA) method for measuring biofilm formation, it was used to determine the persistence of the antibiotic resistance developed by the isolates in biofilms. The antibiotic resistance was found to persist for 3-4 wk post-propagation as planktonic subcultures. Interestingly, some strains even developed resistance to vancomycin and/or teicoplanin. However, no association of either biofilm formation or persistent antibiotic resistance with the major capsular phenotype was observed. These observations highlight the potential significance of (a) determining the antibiograms of S. aureus subcultured from biofilms developed in vitro using the TCP method as well as from planktonic cultures for formulation of an optimal therapeutic strategy, and (b) continuing to identify predominant non-capsular antigens contributing to biofilm formation, regardless of the capsular phenotype for the development of an effective potentially broad-spectrum vaccine for prevention of bovine mastitis caused by S. aureus.

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“…2.2 DNA extraction, amplification, and sequencing of the 16S rDNA gene DNA was extracted as described by Tito et al (2015). The 16S rDNA gene was amplified using the polymerase chain reaction (PCR) technique, using primers 338F 5'-AGAGTTTGATCCTGGCTCAG-3' and 1378R 5'-CGGTGTGTACAAGGCCCGGGAACG-3'.…”

Section: Origin Of the Isolatesmentioning

confidence: 99%

Characterization and evaluation of potential halotolerant phosphate solubilizing bacteria from Salicornia fruticosa rhizosphere

Teles,

Xavier,

Arcênio

et al. 2024

Front. Plant Sci.

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Soil salinization is a significant abiotic factor threatening agricultural production, while the low availability of phosphorus (P) in plants is another worldwide limitation. Approximately 95–99% of the P in soil is unavailable to plants. Phosphate-solubilizing bacteria (PSB) transform insoluble phosphates into soluble forms that plants can utilize. The application of PSB can replace or partially reduce the use of P fertilizers. Therefore, selecting bacteria with high solubilization capacity from extreme environments, such as saline soils, becomes crucial. This study aimed to identify twenty-nine bacterial strains from the rhizosphere of Salicornia fruticosa by sequencing the 16S rDNA gene, evaluate their development in increasing concentrations of NaCl, classify them according to their salinity response, and determine their P solubilization capability. The bacteria were cultivated in nutrient agar medium with NaCl concentrations ranging from 0.5% to 30%. The phosphate solubilization capacity of the bacteria was evaluated in angar and broth National Botanical Research Institute (NBRIP) media supplemented with calcium phosphate (CaHPO4) and aluminum phosphate (AlPO4), and increased with 3% NaCl. All bacterial strains were classified as halotolerant and identified to the genera Bacillus, Enterobacter, Halomonas, Kushneria, Oceanobacillus, Pantoea, Pseudomonas, and Staphylococcus, with only one isolate was not identified. The isolates with the highest ability to solubilize phosphorus from CaHPO4 in the liquid medium were Kushneria sp. (SS102) and Enterobacter sp. (SS186), with 989.53 and 956.37 mg·Kg-1 P content and final pH of 4.1 and 3.9, respectively. For the solubilization of AlPO4, the most effective isolates were Bacillus sp. (SS89) and Oceanobacillus sp. (SS94), which raised soluble P by 61.10 and 45.82 mg·Kg-1 and final pH of 2.9 and 3.6, respectively. These bacteria demonstrated promising results in in vitro P solubilization and can present potential for the development of bioinput. Further analyses, involving different phosphate sources and the composition of produced organic acids, will be conducted to contribute to a comprehensive understanding of their applications in sustainable agriculture.

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Choice of DNA extraction protocols from Gram negative and positive bacteria and directly from the soil

Cited by 8 publications

References 34 publications

Molecular characterization and genetic diversity of Staphylococcus aureus isolates of dairy production farms in Rio de Janeiro, Brazil

Molecular characterization and genetic diversity of Staphylococcus aureus isolates of dairy production farms in Rio de Janeiro, Brazil

The persistence of biofilm-associated antibiotic resistance of Staphylococcus aureus isolated from clinical bovine mastitis cases in Australia

Characterization and evaluation of potential halotolerant phosphate solubilizing bacteria from Salicornia fruticosa rhizosphere

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