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Identification of lactic acid bacteria isolated from vinegar flies and Merlot grapes

Thesis (MSc)--University of Stellenbosch, 2005. / ENGLISH ABSTRACT: Thirty lactic acid bacteria were isolated from the intestinal tract of Drosophila simulans
Stuvervant and nine lactic acid bacteria from Merlot grapes collected from the same
winery in the Stellenbosch region, South Africa.
The isolates were grouped according to morphological, biochemical and physiological
characteristics. Isolates selected from each group were identified to species level by PCR
with species-specific primers, PCR-based DGGE and 16S rDNA sequencing. The
majority of isolates from the intestinal tract of Drosophila simulans Stuvervant belonged
to the species Lactobacillus plantarum, but Lactobacillus paracasei, Lactobacillus
sanfranciscensis, Leuconostoc mesenteroides subsp. mesenteroides, Lactococcus lactis
subsp. lactis, Enterococcus faecalis and Pediococcus pentosaceus were also identified.
As far as we could determine, this is the first report on the isolation of L. paracasei, L.
sanfranciscensis, L. mesenteroides subsp. mesenteroides, L. lactis subsp. lactis, E.
faecalis and P. pentosaceus from vinegar flies. Lactobacillus plantarum has previously
been isolated from Merlot grapes.
The genotypic relatedness among isolates of L. plantarum isolated from the intestinal
tract of vinegar flies and from Merlot grapes were determined by RAPD-PCR. The
isolates were grouped into four genotypically well-separated clusters. Thirteen isolates
from grape must and five from flies yielded identical RAPD-PCR banding patterns and
grouped into one cluster, suggesting that they are descendants from the same strain. This
suggests that L. plantarum has the ability to use vinegar flies as a vector. / AFRIKAANSE OPSOMMING: Dertig melksuurbakterieë is vanuit die dermkanaal van Drosophila simulans Stuvervant
geïsoleer en nege melksuurbakterieë vanuit Merlot-druiwe. Die druiwe is afkomstig van
dieselfde wynkelder in die Stellenbosch-area van Suid-Afrika.
Die isolate is volgens morfologiese, biochemiese en fisiologiese eienskappe gegroepeer.
Verteenwoordigende isolate vanuit die fenotipiese groepe is tot spesievlak met behulp
van lukraak ge-amplifiseerde polimorfe-DNA (RAPD) polimerase ketting-reaksie (PKR),
PKR met spesie-spesifieke inleiers, PKR-gebaseerde denaturerende gradient-jel
elektroforese (DGGE) en 16S rDNA sekwensering geïdentifiseer.
Die meerderheid isolate uit die ingewande van Drosophila simulans Stuvervant is as
Lactobacillus plantarum geklassifiseer. Stamme van Lactobacillus paracasei,
Lactobacillus sanfranciscensis, Leuconostoc mesenteroides subsp. mesenteroides,
Lactococcus lactis subsp. lactis, Enterococcus faecalis en Pediococcus pentosaceus is
ook geïdentifiseer. Sover bekend, is dit die eerste keer dat L. paracasei, L.
sanfranciscensis, L. mesenteroides subsp. mesenteroides, L. lactis subsp. lactis, E.
faecalis en P. pentosaceus uit asynvlieë geïsoleer is. Lactobacillus plantarum is
voorheen uit Merlot-druiwe geïsoleer.
Die genotipiese ooreenkoms tussen die stamme van L. plantarum wat uit die asynvlieë en
Merlot-druiwe geïsoleer is, is deur middel van RAPD-PKR bepaal. Hiervolgens is die
stamme in vier genotipies goed-gedefinieerde groepe geplaas. Dertien isolate vanuit
druiwemos en vyf vanuit asynvlieë het identiese RAPD-PKR bandpatrone vertoon en het
in een groep gesorteer. Hierdie resultate dui daarop dat die stamme heel moontlik uit een
voorouer ontstaan het en dat asynvlieë heel moontlik as vektor vir L. plantarum dien.

Identiferoai:union.ndltd.org:netd.ac.za/oai:union.ndltd.org:sun/oai:scholar.sun.ac.za:10019.1/16606
Date10 1900
CreatorsGroenewald, W. H.
ContributorsDicks, L.M.T., University of Stellenbosch. Faculty of Science. Dept. of Microbiology.
PublisherStellenbosch : University of Stellenbosch
Source SetsSouth African National ETD Portal
Languageen_ZA
Detected LanguageEnglish
TypeThesis
Formatvi, 95 leaves : ill.
RightsUniversity of Stellenbosch

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