Salmonella

Laboratorio de Microbiologa
Aplicada
Dra. Esther Z. Vega Bermdez
Salmonella sp.
 Salmonella

Salmonella Salmonella
enterica bongori

Subs.
enterica
Subs. Subs. Subs.
salamae Subs. Subs.
arizonae diarizonae indica houtenae

From: Porwollik, S., et al., 2004. Characterization of Salmonella enterica subspe

Of Microarrays. J. of Bacteriol. 186:5883-5898.
Incidencia de Salmonella
 Caractersticas

Enterobacterias
Bacilos Gram (-)
Mtiles con
flagelacin pertrica
No esporulan
No fermentan
lactosa
2, 300 especies
Crece entre 40-140oC
 Variantes Genticas
Mediada por Plasmidios
Utilizacin de lactosa y sucrosa
Microorganismos atpicos pueden
escapara la deteccin en situaciones
clnicas
Las especies de Salmonella no son un
grupo homogneo de organismos
 Reservorios / Alimentos
Relacionados
Carnes crudas
Aves
Huevos
Leche y productos lcteos
Pescado
Camarones
Salsas
Mezclas para bizcocho
Postres de crema
otros
 Patognesis

Dosis infecciosa tan bajo como 15-20 clulas

(depediendo de hospedero)
Antgenos estructurales (pared celular o
antgeno O, H- flagelar y el IV-antgeno
capsular termolbil -poder frente a los
cidos)
Sustancias txicas
Endotoxinas
Exotoxinas (efecto irritante y citotxico
para el tracto intestinal)
Ruta oral-fecal
 Sintomatologa y
Enfermedades
Septicemias
Perodo de Bacteremias
incubacin de 6-48hrs Cistitis,
Diarreas
Dolor abdominal
endocarditis y
Fiebre entre 8-72hrs
osteomilitis
Dolor de cabeza y Sndrome de
en el cuerpo Reiters que puede
Escalofros terminar en artritis
Naseas crnica
Vmitos Fiebre tifoidea
paratifoidea
Diagnstico y Tratamiento
Pruebas de serologa y de heces
fecales
Aislar el paciente
Tratamiento para todos en el
hogar
Antiboticos
 Muestreo en Alimentos
Mtodos convencionales de
aislacin que requieren de
cinco das para confirmar
resultados.
Cultivo 37-420C o en nevera.
Salmonella
 Metodologa
Pre-Enriquecimiento
Caldo de lactosa
Permite la recuperacin de clulas heridas
Aumenta la razn Salmonella:Non-Salmonella
Agua de dilucin peptonada
Productos Animales
Dilucin 1:10
I.e.. 25 g muesra a 225 ml de caldo
24 horas a 35 oC
 Enriquecimiento Selectivo
1 ml de caldo de Lactosa
10 ml Selenite Cystine
10 ml tetrathionate broth
Permite el crecimiento de Salmonella
No hay resultados despes de este
paso
24 horas a 35oC
 Placas Selectivas
Tres tipos de medio selectivo
Bismuth Sulfite
Marrn, gris, negro con metallic sheen
Hektoen Enteric Agar
Azul-verdoso con o sin centro negro o glossy black
Xylose Lysine Desoxycholate
Rosado con o sin centro negro
Estriado de Selective Enrichments
Incubacin de 18-24 h a 35oC
Resultados PRESUMPTIVE POSITIVE
 Medio Diferencial
Discernimiento bioqumico
Triple Sugar Iron Agar
Streak Slant and Stab Butt
1% Lactosa
0.1% Glucosa
Sulfato Ferroso
Indicador de H2S
Tiosulfato
Produccin de H2S
Rojo fenol
Alkalina Rojo, Acido Amarillo
Incubacin a 35 oC por 24 horas
 Triple Sugar Iron Agar
Reaccin de Salmonella
Red Slant (Alkalino)
Acid Butt (Amarillo)
Precipitado Negro
Lysine Iron Agar- Discernimiento
bioqumico
Stab Butt and Streak Slant
Se quiere condiciones anaerbicas en el fondo
para que ocurra la decarboxilacin
Decarboxilacin a partir de L-Lysina
Sulfato Ferroso
Bromo Cresol Purple
Violeta Alkalino, Amarillo - Acido
Glucosa
Tiosulfato
 Lysine Iron Agar
Reaccin de Salmonella
Alkaline Butt Purple
Precipitado Negro
Slo es negativo si hay condiciones
cidicas, pueden obtenerse otras
reacciones no tpicas
 Confirmacin Bioqumica
Aglutinacin de antgenos superficiales
con anticuerpos especficos para
Salmonella
Antgenos de Salmonella
O Somtico Lipopolisacrido
H Flagellar Protena
K Capsular - Polisacridos
 Aglutinacin
Mezcle anticuerpos con un poco de
cultivo
Aglutinacin es un resultado positivo
Aislamiento vs. Enumeracin
Usualmente slo se aisla
Se enumera para propsitos de
investigacin
Mtodo MPN
Mtodo de filtracin por membranas
No Accurate
 Otros mtodos- kits
bioqumicos
Table 1. Partial list of miniaturized biochemical kits and automated systems for
identifyingFormat
System foodborneManufactur
bacteria* ( 5, 8, 15, 16, 21, 35, 36 ).
Organisms
APIb biochemi er
bioMerieux Enterobacteriaceae, Listeria, Staphylococcus, Campylobacter,
Cobas IDA cal
biochemi Hoffmann Non-fermenters, anaerobes
Enterobacteriaceae
Micro-ID b cal
biochemi LaRoche
REMEL Enterobacteriaceae, Listeria
Enterotub cal
biochemi Roche Enterobacteriaceae
eII
Spectrum cal
biochemi Austin Enterobacteriaceae
10
RapID cal
biochemi Biological
Innovative Enterobacteriaceae
BBL cal
biochemi Diag.
Becton Enterobacteriaceae, Vibrionaceae, Non-fermenters, anaerobes
Crystal
Minitek cal
biochemi Dickinson
Becton Enterobacteriaceae
Microbact cal
biochemi Dickinson
Microgen Enterobacteriaceae, Gram negatives, Non-fermenters, Listeria
Vitek b cal
biochemi bioMerieux Enterobacteriaceae, Gram negatives, Gram positives
Microlog cal
C a
Biolog Enterobacteriaceae, Gram negatives, Gram positives
MISb oxidation
Fatty Microbial-ID Enterobacteriaceae, Listeria, Bacillus, Staphylococcus,
Walk/Awa acid
a
biochemi
a
MicroScan Campylobacter
Enterobacteriaceae, Listeria, Bacillus, Staphylococcus,
y
Replianaly cal
biochemi
a
Oxoid Campylobacter
Enterobacteriaceae, Listeria, Bacillus, Staphylococcus,
zer
Riboprinte cal
nucleic
a
Qualicon Campylobacter
Salmonella, Staphylococcus, Listeria, Escherichia coli
r
Cobas acida
biochemi Becton Enterobacteriaceae, Gram negatives, Non-fermenters
Micro-IDb
Malthus cala
conducta Dickinson
Malthus Salmonella, Listeria, Campylobacter, E. coli, Pseudomonas,
Bactomet nce
impedan
a
bioMerieux coliforms
Salmonella
*erTable modified
ce from: Feng, P., App.I., FDA Bacteriological Analytical Manual, 8A ed.
a

a
Automated systems
NOTE:
b Thissystems
Selected table is adopted
intendedAOAC
for general
Officialreference onlyAction.
First or Final and lists known available methods. Presence on
this list does not indicate verification, endorsement, or approval by FDA for use in food analysis.
Identificacin basada DNA-DNA

Table 2. Partial list of commercially-available, nucleic acid-based assays used in the

detection of foodborne bacterial pathogens* (2, 5, 8, 12, 14, 22, 25, 36, 37).

Organism Trade Name Format Manufacturer

Salmonella GENE-TRAKc probe GENE-TRAK

BAX PCR Qualicon

BINDb phage BioControl

Probelia PCR BioControl

* Table modified from: Feng, P., App.I, FDA Bacteriological Analytical Manual, 8A ed.
a
Polymerase chain reaction
b
Bacterial Ice Nucleation Diagnostics
c
Adopted AOAC Official First or Final Action
NOTE: This table is intended for general reference only and lists known available methods. Presence
on this list does not indicate verification, endorsement, or approval by FDA for use in food analysis.
Identificacin basada en deteccin
de antgenos
Table 3. Partial list of commercially-available, antibody-based assays for the detection of foodborne pathogens and toxins*
(3, 5, 8, 12, 33, 36).
Organism/toxin Trade Name Assay Formata Manufacturer
Salmonella Bactigen LA Wampole Labs
Spectate LA Rhone-Poulenc
Microscreen LA Mercia
Wellcolex LA Laboratoire Wellcome
Serobact LA REMEL
RAPIDTEST LA Unipath
Dynabeads Ab-beads Dynal
Screen Ab-beads VICAM
CHECKPOINT Ab-blot KPL
1-2 Teste diffusion BioControl
SalmonellaTEKe ELISA Organon Teknika
TECRAe ELISA TECRA
EQUATE ELISA Binax
BacTrace ELISA KPL
LOCATE ELISA Rhone-Poulenc
Assurancee ELISA BioControl
Salmonella ELISA GEM Biomedical
Transia ELISA Transia
Bioline ELISA Bioline
VIDASe ELFAb bioMerieux
OPUS ELISAb TECRA
PATH-STIK Ab-ppt LUMAC
Reveal Ab-ppt Neogen
Clearview Ab-ppt Unipath
UNIQUEe Capture-EIA TECRA
* Table modified from: Feng, P., App.I, FDA Bacteriological Analytical Manual, 8A ed.
a
Abbreviations: ELISA, enzyme linked immunosorbent assay; ELFA, enzyme linked fluorescent assay; RPLA, reverse passive latex
agglutination; LA, latex agglutination; Ab-ppt, immunoprecipitation.
**Automated
b CAUTION:ELISAunless the assays claim that they are specific for the O157:H7 serotype, most of these tests detect only the
O157
c
EHECantigen; hence will E.
- Enterohemorrhagic also react
coli; ETECwith O157 strains
- enterotoxigenic E. that
coli are not of H7 serotype. These O157, non-H7 strains, generally do
NOTE:
not This table
produce is intended
Shiga toxins for general
and are referenceasonly
regarded notand lists known
pathogenic available
for humans. methods. Presencesome
Furthermore, on this list does not
antibodies toindicate
O157 canverification,
also
d
Also detects E. coli LT enterotoxin
endorsement,
cross react or approval
with by FDAE.for
Citrobacter, use in food
hermanii andanalysis.
other enteric organisms.
e
Adopted AOAC Official First or Final Action
Identificacin basada en la utilizacin de
sustratos

Table 4. Partial list of other commercially available rapid methods and specialty
substrate media for detection of foodborne bacteria* (4, 8, 13, 20, 27, 36).
Organism Trade Name Formata Assay Manufacturer

Salmonella Isogridb HGMF QA Labs

OSRT Medium/ motility Unipath (Oxoid)

Rambach Medium CHROMagar

MUCAP C8esterase Biolife

XLT-4 Medium Difco

MSRVb Medium

* Table modified from: Feng, P., App.I, FDA Bacteriological Analytical Manual, 8A ed.
a
Abbreviations: APC, aerobic plate count; HGMF, hydrophobic grid membrane filter; ATP,
adenosine triphosphate; MUG, 4-methylumbelliferyl--D-glucuronide; ONPG, O-nitrophenyl -D-
galactoside; MPN, most probable number.
b
Adopted AOAC Official First or Final Action.
NOTE: This table is intended for general reference only and lists known available methods.
c
Application for water analysis
Presence on this list does not indicate verification, endorsement, or approval by FDA for use in
d
EHEC - enterohemorrhagic Escherichia coli
food analysis.