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Sökning: WFRF:(Vega Alejandro A.) > (2020-2022)

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1.
  • Beal, Jacob, et al. (författare)
  • Robust estimation of bacterial cell count from optical density
  • 2020
  • Ingår i: Communications Biology. - : Springer Science and Business Media LLC. - 2399-3642. ; 3:1
  • Tidskriftsartikel (refereegranskat)abstract
    • Optical density (OD) is widely used to estimate the density of cells in liquid culture, but cannot be compared between instruments without a standardized calibration protocol and is challenging to relate to actual cell count. We address this with an interlaboratory study comparing three simple, low-cost, and highly accessible OD calibration protocols across 244 laboratories, applied to eight strains of constitutive GFP-expressing E. coli. Based on our results, we recommend calibrating OD to estimated cell count using serial dilution of silica microspheres, which produces highly precise calibration (95.5% of residuals <1.2-fold), is easily assessed for quality control, also assesses instrument effective linear range, and can be combined with fluorescence calibration to obtain units of Molecules of Equivalent Fluorescein (MEFL) per cell, allowing direct comparison and data fusion with flow cytometry measurements: in our study, fluorescence per cell measurements showed only a 1.07-fold mean difference between plate reader and flow cytometry data.
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2.
  • Díaz-Sánchez, Adrian A., et al. (författare)
  • Molecular detection and identification of tick-borne pathogens in Equus caballus and ticks from western Cuba : [Detección e identificación molecular de patógenos transmitidos por garrapatas en Equus caballus y garrapatas del occidente de Cuba]
  • 2022
  • Ingår i: Biotecnologia Aplicada. - : Elfos Scientiae. - 0864-4551 .- 1027-2852. ; 39:2, s. 2501-2505
  • Tidskriftsartikel (refereegranskat)abstract
    • Babesia caballi, Theileria equi and several species of rickettsias are agents of vector-borne diseases that affect equines. The objective of this study was to detect infections by B. caballi and T. equi in horses and to identify rickettsias in horses and ticks in the western region of Cuba. Two nPCR assays were developed and standardized for the detection of B. caballi and T. equi. Blood samples from horses and ticks were collected. Identification by blood smear and molecular detection and identification of B. caballi, T. equi and Rickettsia spp. were carried out. Intraerythrocytic formations compatible with B. caballi and T. equi were observed. The nPCR showed that 25 % of the samples were positive for B. caballi, 73 % for T. equi and 20 % showed coinfection. The results were confirmed with the partial sequencing of the genes bc48 (B. caballi) and ema-1 (T. equi). The sequencing of the 18S rRNA gene of T. equi demonstrated the presence of at least two genotypes of T. equi isolates in Cuba. The real time qPCR assay and sequencing revealed the presence of Rickettsia amblyommatis in A. mixtum and Rickettsia felis in D. nitens.Conclusions: These results constitute the first piece of molecular evidence of B. caballi and T. equi in horses and the first report of R. felis in D. nitens in Cuba, which broadens the knowledge about the distribution of pathogens and the spectrum of potential vectors contributing to the strengthening of management and control programs.
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