Αμαλία Δ. Καραγκούνη Καθηγήτρια (Επιβλέπουσα), Αδριανή Πανταζίδου Αναπληρώτρια Καθηγήτρια, Δημήτριος Χατζηνικολάου Επίκουρος Καθηγητής
The presence of microorganisms in pharmaceuticals products, not only threats
consumers’ health but affects negatively the product’s characteristics. The aim
of this study was the investigation of microbial diversity in raw materials and
finished product (sunscreen) of a pharmaceutical industry, using a combination
of culture dependent techniques and molecular methods (genus specific PCR,
BOX-PCR, ΙΤS-PCR, DGGE, 16S rDNA sequencing) and phylogenetic analysis
(GelCompar II, BLAST, MEGA5).
The study was focused towards the purified water, which constitutes the basic
ingredient in the formula of the sunscreen and in particular it was focalized
in the detection and isolation of: a) the heterotrophic and oligotrophic
mesophilic community, b) the psychrophilic community, c) the filterable
bacteria that penetrate the filter membranes with 0.2 μm diameter pores.
The results of this study demonstrate two discrete communities which prevail in
purified water. The heterotrophic community was composed of the genus Bacillus
(dominant species B. aquimaris, B. barbaricus, B. cibi, B. niabensis), whereas
the phylum Actinobacteria (genera Rhodococcus, Kocuria, Microbacterium)
participate lower levels. In oligotrophic community a-Proteobacteria prevailed
(genera Methylobacterium, Blastobacter, Bradyrhizobium, Sphingomonas). The
genus Sphingomonas has been suggested to be the responsible microorganism to
initiate the colonization and formation of biofilms on the surface of the
production units. Later on, other Proteobacteria attach the incipient biofilm.
Other genera identified, were Brevibacillus, Ralstonia, Bulkhoderia,
Acinetobacter and Paracoccus. The use of the DGGE method, revealed similar
fingerprint patterns throughout the samplings. The most noticeable result was
the detection of ribotypes in samples derived from the filtering of membranes
0.2 μm, which suggest the occurrence of VBNC and filterable bacteria.
Concerning the rest of the raw materials examined, two fungi, Penicillium
chrysogenum and Neosartorya hiratsukae were identified in 3 colorants (iron
Finally, sunscreen’s preservatives were very effective against bacteria E. coli
and P. aeruginosa but demonstrated moderate effect against B. subtilis, which
survives and remains detectable after 28 days within the sunscreen. This result
in combination with a) the single strain (B. safensis) which was isolated from
an unopened bottle of sunscreen and b) the wide dispersion in the water the
genus Bacillus, constitutes a factor that needs to be studied more extensively.
Pharmaceutical and Cosmetic Products, Purified Water, Microbial Diversity, Oligotrophic bacteria, Viable but non culturable bacteria (VBNC)