APPLICATION OF PHAGE TYPING AND PULSED-FIELD GEL ELECTROPHORESIS TO ANALYSE SALMONELLA ENTERICA

ISOLATES FROM A SUSPECTED OUTBREAK IN LAGOS, NIGERIA

KABIR O. AKINYEMI ET AL.

María Alejandra Ramírez Ruiz

María Carolina Portela Fernández

UPB - Medellín

INTRODUCTION

Salmonella

Salmonella

Taxonomic classification: Kingdom: Bacteria Class: Gammaproteobacteria Order: Enterobacteriales Family: Enterobacteriaceae Genus: Salmonella

Salmonella enterica

Pulsed-Field Gel Electrophoresis (PFGE)

Used to analyze molecules separates them according to their size & molecular weight.

Uses electric flow to ease migration.

ONE OF THE MOST USED TECHNIQUES IN

MOLECULAR BIOLOGY RESEARCH

Relationship in between

GENERAL OBJECTIVE

General objective

To identify the prevalences of the different Salmonella enterica serovars using phagotyping and PFGE techniques in Lagos, Nigeria.

MATERIALES Y MÉTODOS

Bacterias

Condiciones óptimas

de conservaci

ón

* Las muestras eran de S. enteritidis y/o de S. typhimurium.

Test de Susceptibilidad Bacteriana

Test de Susceptibilidad Bacteriana

Susceptibilidad a 13 antibióticos fue evaluada:

Ampicilina, amoxicilina, gentamicina, tetraciclina, cloramfenicol, sulfato de colecistina, trimetoprim-sulfometoxazol, trimetoprim, cefoperazona, ofloxacina, ciprofloxacina, perfloxacina y cefotaxima.

Controles: Escherichia coli ATCC 25922 Staphylococcus aureus ATCC 29213 Enterococcus faecalis ATCC 29212

Fagotipificación

Usa bacteriófagos

ID serotipos comunes

Mismo fagotipo Relación epidemiológica

Electroforesis en gel de campo pulsado (PFGE)

Ha demostrado ser altamente discriminatoria en epidemiología

RESULTADOS

Salmonella tymphimurium

Salmonella tymphimurium

PFGE: X1 y X2: 96.8% similitud grupo 1. X3: mayor prevalencia (57.9%) grupo 2. X4 grupo 3.

Fagotipos Grupo 1: RDNC Grupo 2: DT71 Grupo 3: UT Total: 19 muestras

Similitud total: 71%

Salmonella tymphimurium

SEROTIPO UBICACIÓN

FAGOTIPIFICACIÓN

PFGE CANTIDAD

Salmonellatyphimuriu

m

Humanos RDNC X1 2

Humanos RDNC X2 2

Agua RDNC X2 1

Humanos DT71 X3 5

Ganado DT71 X3 2

Agua DT71 X3 2

Aguas sucias

DT71 X3 2

Humanos UT X4 3

TOTAL: 19RDNC: Reaccionaron pero no se conformaronDT71: Reaccionaron y se conformaron con ese fago.UT: No se tipificaron

Salmonella enteritidis

Salmonella enteritidis

PFGE X1e: más frecuente (61.5%) grupo 1.

MULTIRRESISTENCIA X2e y X3e: similitud 80% grupo 2. X4e grupo 3.

Fagotipos PT29 UT RDNC

Total: 13 muestrasSimilitud total: 64%

Salmonella enteritidis

SEROTIPO

UBICACIÓN

FAGOTIPIFICACIÓN

PFGE CANTIDAD

Salmonellaenteritidis

Humanos UT X1e 4

Aguas residuales

UT X1e 1

Humanos RDNC X1e 3

Humanos RDNC X2e 1

Humanos RDNC X3e 1

Agua RDNC X4e 1

Ganado RDNC X4e 1

Agua PT29 X4e 1

TOTAL: 13RDNC: Reaccionaron pero no se conformaronPT29: Reaccionaron y se conformaron con ese fago.UT: No se tipificaron

DISCUSSION

AUTHOR STATEMENT YES NO

Laconcha I et al. PFGE and phagetyping increase the discrimination in the

epidemiologic analysis of S. enteriditis strains.

X

Onyango MD et al. Phagetyping profiles cathegorized as 1, 3 & 5 may be found in S.

typhimurium in AfricaX

Ward LR et al. Phage type PT4 is the most found within S. enteritidis samples.

X

Porwollik S et al. Serogroup, host specificity & disease characteristcs influence

the isolates’ clustering.X

CONCLUSIONS

Conclussions

The characterization and research about microorganisms as the Salmonella is absolutely necessary to improve the control of the diseases they cause.

It is alarming to see how the bacteria are getting each day stronger against antibiotics (out of 13 antibiotics just 3 showed to be effective against all serovars) and that’s mainly led by the self-medication with antibiotics, not following correct treatment schemes.

Conclussions

The identification of different Salmonella serotypes can help to establish control on the transmission mechanisms used by these microorganisms generating a positive impact on the community’s public health.

Thanks to such studies we can getsome control over the use of antibiotics and try to educate people about their use and how NOT to use them, to prevent multidrug resistance.

BIBLIOGRAFÍA

Akinyemi KO, Phillip W, Beyer W, Böhm R. Application of phage typing and pulsed-field gel electrophoresis to analyse Salmonella enterica isolates from a suspected outbreak in Lagos, Nigeria. J Infect Dev Ctries 2010; 4 (12): 828-834.

Mancera-Martínez A, Vázquez Navarrete J, Heneidi-Zeckua A. Phage typing of Salmonella enteritidis strains isolated from poultry in Mexico. Téc Pecu Méx 2004; 42 (2): 287 – 294.

MARTINEZ SÁNCHEZ, Lina María [y otros]. Biología molecular. 6 ed. Medellín: UPB. Fac. de Medicina, 2011. 292 p.

Autor desconocido.“Pruebas de sensibilidad a agentes antimicrobianos”, disponible online en http://minnie.uab.es/~veteri/21273/Practica%202.2009-10.pdf [consultado 24/02/2011, 16:30 hrs]

Autor desconocido.“Prueba de susceptibilidad antimicrobiana por difusión en agar”, disponible online en http://www.ispch.cl/lab_sal/doc/man_suscep.pdf [consultado 24/02/2011, 19:00 hrs].

MAPAS CONCEPTUALES

Por: María Carolina Portela Fernández

Por: María Alejandra Ramírez Ruiz