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Molecular investigation of macrolide and Tetracycline
resistances in oral bacteria isolated from Tunisian children
Tarek Zmantar et ál
Steffany Franco SañudoCamilo Díaz Quintero
Medical SchoolMolecular Biology
III SemesterUPB2011
MACROLIDES Are a group of closely related
compounds characterized by a macrocyclic lactone ring to which deoxysugars are attached that have antimicrobial activity
Erythromycin Prototype drug
Katzung Bertram, Farmacología básica y clínica. 11ª ed. Mc graw Hill
Antimicrobial activity
Against gram-positive and negative organisms
Inhibitory Bactericidal
Reducing the protein synthesis
In high concentrations
Acting like
Can Produce
Resistence
Reducing permeability of the
cell membrane
Productionof esterases
Modification of the ribosomal binding site
by
Katzung Bertram, Farmacología básica y clínica. 11ª ed. Mc graw Hill
TETRACYCLINES
Are broad-spectrum bacteriostatic antibiotics that inhibit protein synthesis.
Doxycycline Example drug
Katzung Bertram, Farmacología básica y clínica. 11ª ed. Mc graw Hill
Antimicrobial activity
Against gram-positive and negative organisms
Passive difussion Active process
Reducing the protein synthesis
Acting like
Can Produce Resistence
Decreased intracellular
accumulation
Ribosome protection
Enzymatic inactivation of tetracyclines
by
Inhibitoryby
DRUG RESISTENCEA decrease in sensivity to suficient degree to by reasoneably certain that a strain concerned is different from a sample of wild stains of human type that have never come in contact with the drugs.
A decrease in sensivity to suficient degree to by reasoneably certain that a strain concerned is different from a sample of wild stains of human type that have never come in contact with the drugs.
Katzung Bertram, Farmacología básica y clínica. 11ª ed. Mc graw Hill
SUSCEPTIBILITYAn organism is called
“susceptible” to a drug when the infection caused by it is likely to respond to treatment with this
drug, at the recommended dosage.
An organism is called “susceptible” to a drug when the infection caused by it is likely to respond to treatment with this
drug, at the recommended dosage.
Katzung Bertram, Farmacología básica y clínica. 11ª ed. Mc graw Hill
GENERAL OBJETIVE
Determine the prevalence of antibiotic
susceptibility and the distribution of
erythromycin resistance gene (ermB), the
macrolide efflux (mefI) and tetracycline
resistance genes (tetM, tetO) in oral bacteria
including streptococci and enterococci
isolated from Tunisian children.
Determine the prevalence of antibiotic
susceptibility and the distribution of
erythromycin resistance gene (ermB), the
macrolide efflux (mefI) and tetracycline
resistance genes (tetM, tetO) in oral bacteria
including streptococci and enterococci
isolated from Tunisian children.
PACIENTES
CEPAS• Muestras extraídas de la cavidad oral
de los pacientes
• Se escogieron placas dentales y caries
• Incubadas en medios BHI con sangre
de oveja desfibrinada al 5%
• Se lograron identificar
ESTREPTOCOCOS con tiras de API20
• Se analizaron los resultados con un
mini API
PRUEBAS DE SUSCEPTIBILIDAD
Pruebas de ATB Strep Strip
Es una prueba de susceptibilidad a los antibióticos para el estreptococo que
contiene un rango de 18 tipos diferentes fármacos.
Pruebas de ATB Strep Strip
Es una prueba de susceptibilidad a los antibióticos para el estreptococo que
contiene un rango de 18 tipos diferentes fármacos.
DETECTION DE GENOTIPOS DE RESISTENCIA POR PCR
CARACTERIZACIÓN BIOQUÍMICA
• Se aislaron e identificaron 109 cepas por API 20
• Más aislada Streptococcus mutans
• Se identificaron 15 cepas relacionadas con la aparición de caries
SUSCEPTIBILIDAD ANTIBIÓTICA
De las 109 cepas aisladas se identificó
resistencia a la estreptomicina en un 22% correspondiente
a 24 cepas.
De las 109 cepas aisladas se identificó
resistencia a la estreptomicina en un 22% correspondiente
a 24 cepas.
SUSCEPTIBILIDAD ANTIBIÓTICA
La susceptibilidad intermedia fue del
96.3% para la Kanamicina y la
Gentamicina.
La susceptibilidad intermedia fue del
96.3% para la Kanamicina y la
Gentamicina.
Todas las cepas fueron susceptibles para la
Ampicilina Enterococcus, Telitromicina y Teicoplanina.
Todas las cepas fueron susceptibles para la
Ampicilina Enterococcus, Telitromicina y Teicoplanina.
SUSCEPTIBILIDAD ANTIBIÓTICA
DETECCIÓN DE LOS GENES DE RESISTENCIA POR PCR
Electroforesis en gel de agarosa de PCR multiple amplificando los genes ermB (639 pb) y mefI (263 pb).
DETECCIÓN DE LOS GENES DE RESISTENCIA POR PCR
Electroforesis en gel de agarosa de PCR convencional amplificando el gen tetM (1862 pb).
Electroforesis en gel de agarosa de PCR convencional amplificando gen tetO (538 pb).
DETECCIÓN DE LOS GENES DE RESISTENCIA POR PCR
Author Plot Agree or
Disagree
Warren JJ et ál.
Dental caries is closely associated with mutans streptococci colonizing the oral cavity.
Villedieu A et ál.
Tetracycline-resistant streptococciare frequently isolated from the oral cavity of humans
Author Plot Agree or
Disagree
Martel A et ál.
Acquired resistance against these antibiotics has frequently been described in enterococci.
Lockhart PB et ál.
The failure of antibiotic prophylaxis in dental practice is the major cause of bacteremia after dental procedures. The ability of antibiotic therapy to reduce the frequency of bacteremia associated with dental procedure is controversial
• This investigation can be applied to the daily
procedures because it’s relevant to prevent sistemics
infections.
• The study demostrate the importance of pair differents
subjects such as dentistry, genetic and medicine all for
the patient wellbeing.
• Cariogenic diet and oral hygiene is crucial for the
contribution of the non-development of caries, also the
failure of antibiotic prophylaxis in dental practice is the
major cause of bacteremia after dental procedures.
• Isolated strains showed antibiotic susceptibility of
tetracycline that can be detected by PCR of resistence
genes has frequently been described in
enterococci.
Camilo Andrés Díaz Quintero
Steffany Franco Sañudo