PCR-DGGE assessment of the bacterial diversity of breast milk in women with lactational infectious mastitis

<p>Abstract</p> <p>Background</p> <p>Infectious mastitis is a common condition during lactation and in fact, represents one of the main causes leading to a precocious weaning. The number of studies dealing with lactational mastitis is low and, up to now, the etiological diagnosis is frequently made on the basis of unspecific clinical signs. The aim of this study was to investigate the microbial diversity of breast milk in 20 women with lactational mastitis employing culture-dependent and culture-independent (PCR-DGGE) approaches.</p> <p>Methods</p> <p>Breast milk samples were cultured in different media to investigate the presence of bacteria and/or yeasts, and a total of 149 representative isolates were identified to the species level by 16S rRNA gene PCR sequencing. The microorganisms recovered were compared with those found by PCR-DGGE analysis. To identify the DGGE profiles two reference markers of different microbial species were constructed. Sequence analysis of unknown bands was also performed.</p> <p>Results</p> <p>Staphylococci were the dominant bacterial group and <it>Staphylococcus epidermidis </it>was the dominant species. In a lower number of samples, other bacteria (mainly streptococci and a few gram-negative species) were also identified. Globally, PCR-DGGE results showed a good correlation with those obtained by culture-based methods. However, although DNA bands corresponding to different lactic acid bacteria were detected, such bacteria could not be isolated from the milk samples.</p> <p>Conclusion</p> <p>Staphylococci seem to be the main etiological agents of human lactational mastitis. The combined use of culture and molecular techniques allowed a better characterization of the bacterial diversity in milk from women suffering from infectious mastitis. Our results suggest that this condition could be the result of a disbiotic process where some of the bacterial species usually present in human milk outgrow (staphylococci) while others disappear (lactobacilli or lactococci).</p

CIF applications. XVI. CIF2CRY: for CIF input into the CRYSTALS program

CIF2CRY is a program for converting structural data and reflection data contained in a CIF into instructions to be read by the CRYSTALS program [Betteridge, Carruthers, Cooper, Prout and Watkin (2003). J. Appl. Cryst. 36, 1487]. The program can be used, for example, to read in CIFs that are archived by crystallographic journals, allowing users to study and re-refine published structures. Multiple data blocks in one CIF can be converted into a continuous file of CRYSTALS instructions, including an instruction for running user-specified calculations after each structure has been input. The program uses the CIFtbx Fortran library