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This article was downloaded by [Southern Illinois University]On 18 July 2015 At 2105Publisher Taylor amp FrancisInforma Ltd Registered in England and Wales Registered Number1072954 Registered office 5 Howick Place London SW1P 1WG

Journal of Environmental

Science and Health

Part A ToxicHazardous

Substances and

EnvironmentalEngineeringPublication details including instructionsfor authors and subscription informationhttpwwwtandfonlinecomloilesa20

Quantitative

determination of Ecoli and fecal coliforms

in water using a

chromogenic mediumJL Alonso

a A Soriano

b I Amoros

a amp

MA Ferrusc

a Instituto de Hidrologiacutea y Medio Natural Universidad Politeacutecnica Camino de Vera14 Valencia 46022b Gamaser SL c Pedrapiquers 4

Valencia 46014c Departamento de Biotecnologiacutea

Universidad Politecnica SpainPublished online 15 Dec 2008

To cite this article JL Alonso A Soriano I Amoros amp MA Ferrus (1998)Quantitative determination of E coli and fecal coliforms in water using achromogenic medium Journal of Environmental Science and Health Part AToxicHazardous Substances and Environmental Engineering 336 1229-1248DOI 10108010934529809376785

To link to this article httpdxdoiorg10108010934529809376785



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D o w n l o a d e d b y [

S o u t h e r n I l l i n o i s U n i v e r s i t y ] a t 2 1 0 5 1 8 J u l y 2 0 1

5



J EN VIRON SCI HE ALTH A33(6) 12299830851248 (1998)

QU NTIT TIVE DETERMIN TION OF E COLI ND FEC L

COLIFORMS IN

W TER

USING CHROMOGENIC

M E I U M

Key Words Kcoli β 983085 galactosidase

β 983085 glucuronidase

water fecal coliforms

J L Alonso

1

A Soriano

2

I

Amoros

1

and MA

F e r rus

3

1

Inst i tu to

de Hidrologiacutea y Medio Natural Universidad Politeacutecnica Camino de Vera 14

46022 Valencia

2

Gamaser SL c Pedrapiquers 4 46014 Valencia

3

Departamento de Biotecnologiacutea Universidad Politecnica

Spain

STR CT

A new medium Chromocult Coliformreg Agar (CC agar) developed by E

Merck AG (Darmstadt Germany) was compared with the Standard Methods

membrane filtration fecal coliform (mFC) medium for fecal coliform detection and

enumeration In the CC agar non-E coli fecal coliforms (Klebsiella Enterobacter and

Citrobacter) (KEC) were identified by the production of a salmon to red colour from

p-galactosidase (LAC) cleavage of the substrate Salmon-GAL while E coli colonies

were detected by the blue colour produced by the cleavage of X-glucuronide by

β 983085

1229

Copyright copy 1998 by Marcel Dekker Inc wwwdekkercom

D o w n l o a d e d b y [ S o u t h e r n I l l i n o i s U n i v e r s

i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5



1230 ALONSO ET AL

glucuronidase

G U S ) Statistically there was no significant differences between fecal

coliform counts obtained with the two media (CC agar and mFC agar) and two

incubation

procedures (2h983085 37degC plus 22h983085 445deg and 445degC) as determined by variance

analysis In our study K

coli

represented on average 705983085 925 of the fecal coliform

population A high incidence of

false

negative KEC (195 ) and

E

coli (296 )

colonies

was detected at 445degC Two

K

coli

GUS negative phenotype upon

reinoculation into CC agar were

G U S

+

A total of 31 KEC LAC colonies were

streaked on to

CC

agar and incubated at 37degC 29 KEC strains tha t failed to produce β 983085

galactosidase at 445degC were able to produce the enzyme at 37degC In our opinion the

physiological condition of the fecal coliform isolates could be responsible for the non983085

expression of P983085 galactosidase and P983085 glucuronidase activities at 445degC

INTRODUCTION

The

detection and enumeration of indicator organisms are of primary

importance

for the monitoring of sanitary and microbiological quality of water Fecal

coliforms have been long used as indicators of fecal contamination in water and food

The

term fecal coliform include all coliforms

tha t

can ferment lactose at 445degC trying

t o separate the non ubiquitous coliforms from those of true fecal origin (Dockins and

McFe t e r s 1978) The presence ofpound coli directly relates to fecal contamination with its

implied threat of the presence of enteric disease agents (Rice et al 1990) The other

members of the fecal coliform group Klebsiella Enterobacter and

Citrobacter)

may

be isolated in feces but their presence does not always suggest fecal contamination

D o w n l o a d e d b y [ S o

u t h e r n I l l i n o i s U n i v e r s i t y ] a t 2 1 0 5 1 8 J u l y 2 0 1 5



QUANTITATIVE DETERMINATION OF E COLI 1231

(Covert et al 1992) The abbreviation KE C will be used in this study for the

designation of non-E coli fecal coliforms (Klebsiella Enterobacter and Citrobacter)

A major limitation of current membrane filtration methods used for counting

fecal coliforms is the enumeration of microorganisms which are not exclusively of fecal

origin thereby giving a false indication of the sanitary quality of the water

(Au goustinos et al 1993) Identification of pound coli in the past has been laborious and

only recently methods have been developed that detect

E coli

rapidly with accuracy

and specificity (Alonso et al 1996 Shadix et al 1993) The identification of coliforms

based on detection of P-galactosidase activity (Manafi et al 1991) is a significant

departure from methods that utilize the bacterial end products of lactose fermentation

(APHA 1995)

A new chromogenic medium Chromocult Coliformreg agar (CC agar) has been

developed by E Merck AG (Darmstadt Germany) to detect coliforms and K coli

simultaneously A combination o f tw o chrom ogenic glycosides is used for th e detection

of p-galactosidase (LAC) and P-glucuronidase (GUS) The Salmon-GAL substrate

causes a salmon to red colour of the KEC coliform colonies (LAC

+

GUS ) and the

substrate X-glucuronide is used for the identification of P-glucuronidase E coli

cleaves both Salmon-GAL and X-glucuronide so that positive colonies take on a dark

blue to violet colour (LA C

+

GUS)

In this study the CC agar was compared w ith the M-FC medium recommended

in Standard Methods (1995) for the enumeration of fecal coliforms by the membrane

filtration technique


u t h e r n I l l i n o i s U n i v e r s

i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5



1232 ALONSO ET AL

M A T E R I A L A N D M E T HOD S

Sampling

A total o f 40 w ater samples were collected from 6 different environmental

sources in the Valencia area The water samples were as follows 6 samples from the

Turia river near Valencia drinking water treatment plant (site TR) 11 samples from

two well water supplies (site PI 6 samples and site P2 5 samples) 8 samples from a

heavily polluted stream (site AP) 8 samples of seawater (salinity 21o) (Malvarrosa

beac h) influenced by sew age discharge (site M l ) and 7 samples of seawater from a

point located 200 m south of the previously mentioned sewage discharge (salinity

34o) (site M 2) All samples we re collected in sterile glass bo ttles refrigerated and

assayed within 24 h after collection Samples from sites TR A P M l and M 2 w ere

preassayed within 2 h to estimate bacterial density Several dilutions of these samples

were filtered to estimate the number of KEC and

K coli

present in collected waters

After 22 h incubation the most appropriate dilution was chosen and samples were

definitively analyzed

Bacterial Strains

Th e 32 reference strains from the Coleccioacuten Espantildeola de Cultivos Tipo (C EC T)

and 6 Salmonella strains from the Instituto de Hidrologiacutea y Medio Natural (IHMN)

stock culture collection used in this study are listed in Table 1 All strains except

Enterococcus strains were grow n and maintained on nutrient agar (M erck ) Th e

Enterococcus strains were grow n and maintained on brain heart agar (M erck)

Incubation Temperature Effect

Pure-culture studies were conducted with reference and IHMN strains

Bac teria we re resuspended in 5 ml of phosph ate buffer (APH A 1995) A loopfiil of th e





QUANTITATIVE DETERMINATION OF E OLI

TABLE 1

Growth Conditions a t 37deg 41deg and 44 5degC of Different

Bacteria on Chromocult Coliform Agar

1233

Test Strain

Enterobacter aerogenes

Enterobacter cloacae

Enterobacter sakazakii

Enterobacter gergoviae

Klebsiella pneumoniae

Klebsiella oxytoca

Klebsiella ozaenae

Citrobacter diversus

Citrobacter amalonaticus

Citrobacter freundii

Escherichia coli

Hafnia alvei

Serratia odoriacutefera

Serratia marcescens

Serratia rubiadea

Cedecea davisae

Kluyvera ascorbata

Shigella flexneri

Shigella boydii

Shigella sonnei

Aeromonas hydrofila

Aeromonas caviae

Aeromonas media

A eromonas jandaei

Aeromonas schubertii

Aeromonas trota

Aeromonas eucrenophila

A

veronii bv veronii

Vibrio cholerae

Pseudomonas aeruginosa

Enterococcus faecalis

Enterococcusfaecium

Salmonella derby

S bredeney (4 strains)

Salmonella london

No

684

194

858

857

140

860

851

856

863

401

678

157

867

159

868

842

861

585

583

413

398

838

4232

4228

4241

4255

4224

4257

557

108

184

410

IHMN

IHMN

IHMN

37degC

G

b

C

c

+ r

+ r

+ r

+ r

+ r

+ r

+ r

+ r

+ r

+ r

+ b

+ r

+ r

+ r

+ r

+ c

+ r

+ c

+ t

+ b

+ r

+ r

+ r

+ r

+ c

+ r

+ r

+ r

+ r

+ c

-

-

+ c

+ t

+ t

41

G ~

+

+

+

+

+

+

+

+

+

+

+

+

+

+

-

+

+

+

+

+

+

+

+

-

+

+

+

+

+

-

-

+

+

+

degC

r

r

r

r

r

r

r

r

c

r

b

r

r

t

r

-

r

c

t

b

r

r

r

r

-

r

r

r

r

c

-

-

c

t

t

445degC

G C

+ r

+ r

+ r

-

+ c

+ r

+ r

+ c

+ c

+ r

+ b

+ r

+ r

+ r

+ r

-

+ r

+ c

+ t

+ b

-

-

+ r

-

-

-

-

-

+ c

-

-

+ c

+ t

+ t

No of reference strain from the CECT

G

Growth +=Good +=Weak -=None

b

C

Colour r=Salmon to Red b=Dark Blue to Violet t=Light Blue to Turquoise

c=Colourless





1234 ALONSO ET AL

phosphate buffer culture was streaked onto CC agar plates and incubated at three

different incubation temperatures (37degC 41degC and 445degC) After growth was

observed the P-galactosidase and p-glucuronidase activities of 32 reference strains and

6 Salmonella strains w ere tested

Microbiological Analysis

Samples were decimal diluted or concentrated according to the expected

bacterial density as above described Duplicates of each sample dilution were filtered

through sterile 045 urn pore size membranes (Whatman) using the standard membrane

filtration technique The membranes were placed onto a pre-prepared layer of CC agar

in a 47-mm petri-dish These were then incubated at 445degC in a water bath for 24 h

All salmon to red colonies (LAC

+

GUS ) were counted as presumptive KEC coliforms

and all blue to violet colonies (LAC GUS) were counted as presumptive

K coli

For

comparison the second duplicate membrane of each pair was processed by a standard

method for fecal coliforms The membranes were layered onto M-FC agar (Merck) and

incubated at 445degC in a water bath for 24 h All blue colonies were counted as fecal

coliforms (APHA 1995) Rosolic acid from M-FC medium was eliminated as

suggested by Presswood and Strong (1978) These authors observed that eliminating

rosolic acid from M-FC medium improves the M-FC procedure by allowing higher

fecal coliform colony recoveries

In the modified method the membranes were placed on CC agar and M-FC

agar and were incubated at 37degC for 2 h before incubation at 445degC in a water bath

for 22-2 4 h Ro se et al (1975) suggested the need for a repair phase prior to incubation

at the elevated temperature



i t y ] a t 2 1 0 5 1 8 J u l y 2 0 1 5




A total number of 587 colonies from the most appropriate dilution of CC agar

were submitted to qualitative analysis For each sample site salmon to red colonies

(LAC

+

GUSO dark blue to violet colonies (LAC GUS

4

) light blue to tu rquo ise (LAC

GUS) and colourless colonies (LAC GUS ) were randomly picked and subcultured on

nutrient agar (Merck) Purified cultures were further identified by the following cultural

characteristics indole production growth on Simmons citrate agar (Merck) methyl

red and Voges-Proskauer reactions gas production in EC broth (Merck) reaction on

triple sugar iron agar (TSI) (Merck) and possesion of cytochrome oxidase and

catalase A total number o f 66 isolates were further identified using the A PI 20 E

system (bioMerieux)

Statistical Analysis

Bacterial counts were logarithmically transformed prior to statistical treatment

Results were analyzed by linear regression to verify the linearity of the relationship

between E coli and KEC coliforms obtained with CC agar To examine the medium

performance (CC agar) over a range of sample types and concentrations the samples

w ere groupe d by sample site by E coli and KEC coliform counts on CC agar by fecal

coliform counts on mFC agar and by incubation temperatures A unifactorial variance

analysis was performed on the means of the data All statistics were obtained using

Statgraphics software

RESULTS AND DISCUSSION

E coli and K E C cou nts on CC agar and fecal coliform coun ts on mF C aga r at

two incubation procedures are compared in Table 2 In this study E coli was isolated



i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5



1236 ALONSO ET AL

TABLE 2

N o n - pound

coli

Fecal Coliforms

Klebsiella

spp

Enterobacter

spp and

Citrobacter

spp) KE C ) and

Escherich ia coli

Recovered on Ch romocult Coliform Ag ar

(C C aga r) and Fecal Coliform s Recovered on M FC Agar

Sampling

source

TR

EC-CCA

b

KEC-CCA

C

EKEC-CCA

d

F C - m F C

PI

EC-CCA

KEC-CCA

EKEC-CCA

FC-mFC

AP

E C - C C A

K E C - C C A

EKEC-CCA

FC-mFC

M l

EC-CCA

K E C - C C A

EKEC-CCA

FC-mFC

M2

EC-CCA

K E C - C C A

E K E C - C C A

FC-mFC

Mean

215

197

239

245

195

081

150

168

672

596

679

673

535

451

541

536

307

253

318

313

2h37deg-445degC

SD

151

163

156

167

062

025

080

071

020

014

019

019

124

112

122

125

093

083

091

098

Min

070

030

085

085

111

048

048

060

646

578

656

652

338

270

346

336

208

154

220

208

Max

408

420

445

461

258

108

259

262

700

623

705

703

672

570

676

671

448

370

454

462

Mean

287

264

237

241

179

060

161

161

671

582

676

673

532

439

537

536

300

240

311

307

445

SD

141

148

157

158

055

026

072

077

019

016

018

019

122

129

123

123

102

085

098

101

degC

Min

158

100

078

104

118

030

048

048

651

560

657

652

338

230

341

338

194

140

205

181

Max

426

420

453

448

250

095

250

256

699

611

703

704

669

604

678

668

457

378

463

458

Data are reported as log values per 100 ml The results are expressed as

arithmetic mean (M ean) standard deviation (SD ) minimum (Min)

and maximum (Max)

EC-CCA = Escherichia coli (LAC GUS) recovered on C C agar

K E C-C C A = N o n - pound coli fecal coliforms (LAC

+

GUS ) recovered on CC agar

d

EKEC-CCA = E coli and non-Ecoli fecal coliforms recove red on CC agar

T C -m F C = Fecal coliforms recovered o n mFC agar


i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5




from all of the six zones analyzed but at different densities (Table 2) The data of site

P2 were not reported because of low number of samplings with positive results The

highest levels of E coli w ere detected at sites AP and M l with densities up to 10

s

CFU100 ml These zones also showed high numbers of KEC coliforms Table 3

summarizes the values of the correlation coefficients (r) and the confidence levels (P)

obtained b etween th e concentrations of K coli and KE C At site P2 the presence of E

coli (1 CFU100 ml) was detected only in four samples and it was not included in the

statistical analysis Positive correlations (Plt001) were found at sites TR M l and M 2

Th ere was no correlation at sites P I and AP Counts of E coli and KEC on CC agar

were compared with fecal coliform counts on mFC agar Statistically there was no

significant differences between coliform counts obtained with the two media (CC agar

and mF C agar) and two incubation procedures (2h-37degC plus 22h-445degC and 4 45degC )

as determined by variance analysis ANOVA on the K coli data at two incubation

procedures of CC agar indicated no significant differences among incubation

proc edu res K EC coliforms represented on average 79-2 95 of the fecal coliform

popu lation Figueras et al (1994 ) demonstrated the low specificity o f mF C m edium for

the enum eration and detection of fecal coliforms from seawater on th e basis of the high

incidence of false positive colonies (thermotolerant non-fecal coliforms) Many authors

(Caplenas and Ka narek 1984 Charriere et al 1992 Dufour 1977 Evison 1988)

consider tha t the adjective fecal is no t properly applied and questioned the usefulness

of fecal coliforms other than E coli as fecal indicators We agree with other authors

(Brodsky 1997 Mossel 1997) that in order to provide more comparative results the





1238

ALONSO ET AL

T A B L E 3

Regression an d Correlation Parameters from Data O btained Us ing Chrom ocu lt

Coli form Agar CC Agar)

Sample

site

T R

P I

AP

M l

M 2

Parameters

EC37-KEC37

EC44-KEC44

EC37-KEC37

EC44-KEC44

EC37-KEC37

EC44-KEC44

EC37-KEC37

EC44-KEC44

EC37-KEC37

EC44-KEC44

R

099

098

069

064

068

040

099

099

099

098

P

lt001

lt001

N S

b

N S

N S

N S

lt001

lt001

lt001

lt001

Intercept

(a)

0340

0405

0571

1729

0977

3981

0419

1185

0257

0170

Slope

(b )

0916

0936

1701

0108

0963

0468

1093

0941

1112

0182

a

EC37-FC37=poundscOTc7ij coli and non-pound coli fecal coliforms (Klebsiella

Enterobacier and Citrobacter) (KEC) recovered on CC agar (2h 37deg-445degC) EC44-

FC44=pound coli and non-pound coli fecal coliforms recovered on CC agar (445deg C)

^ 5 = ^ 1 significant

term fecal coliform should be revised and replaced with the more definitive fecal index

organism Escherichia coli

The p-galactosidase and P-glucuronidase activities of 32 reference strains and 6

Salmonella strains at 37degC 41degC and 445degC are show n in Table 1 Th e ability to

produce p-galactosidase of Klebsiella pneumoniae Citrobacter diversus and C

amalonaticus strains on CC agar was inhibited at 445degC The grow th o f Aeromonas

reference strains was inhibited at 445degC except in the case of

Aeromonas jandaei


i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5




Salmonella bredeney (4 strains) and S london showed P-glucuronidase activity at the

three temperatures tested

The identities of the four types of colonies (LAC

+

G U S L A C

+

G U S

+

LAC

GUS and L A C GUS ) on C C agar are shown in Table 4 The identity of 66 isolates

was verified with the API 20E system (Table 5) The KEC LAC

+

GUD species

identified were Klebsiella oxytoca (2 strains) K pneumoniae (2 strains) Enierobader

cloacae (4 strains) Citrobacterfreundii (6 strains) and C amalona ticus (1 strain)

Of the 212 blue colonies (LAC

+

GUS

4

) 207 (98 ) were confirmed as E coli

giving a false positive rate of 2 (5 o f 212 colonies) A total of 9 L A C G U S colonies

15 L A C G U S

+

colonies and 8 7 LA C

+

G US were E coli resulting in a false negative

rate of 296 (111 of 375 colonies) Covert et al (1992) reported that the false-

negative rates with natural populations of E coli ranged from 186 with the

Coliquikreg test (C L) t o 23 4 with the Colilertreg test (C L) (these enzyme detection tests

contains the fluorogenic substrate 4-methylumbelliferyl-P-D-glucuronide M U G )

Ciebin et al (199 5) enco untered a lower incidence of P-glucuronidase-negative E Coli

isolates with river (98 and 93) and lake (78 and 88) samples with FC-BCIG and

TEC-BCIG media (m-FC and m-TEC media supplemented with the chromogenic

substrate 5-bromo-6-chloro-3-indolyl-P-D-gIucuronide BCIG) respectively Two E

coli GUS negative phenotype at 445degC were incubated on CC agar at 37degC to

determine whether the expression of GU S formation was temperature dependent Bo th

E coli strains showed GUS production at 37degC Alonso et al (1996) found that false

negative K coli G U S colonies occurred less frequently at 35degC than at 445 degC S everal

auth ors (C lark et al 1 991 Cov ert et al 1992 Palmer et al 1995) showed that som e

M U G negative Ecoli isolates regained the M U G phenotype upo n further culture O ne

mechanism that could cause GUS negative phenotype would be failure of the permease


i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5



24

ALONSO ET AL

TABLE 4

Num ber o f

coli

a nd N o n-E

coli

Fecal Coliforms Isolates G row n on C C Aga r

Identified on the Basis of IMV IC Cytochrom e O xidase Catalase and TSI Ag ar

Reactions

Phenotype

LAC GUS

AP

C

M l

M 2

TR

PI

P2

Total

LAC

+

GUS

+

AP

M l

M 2

T R

PI

P2

Total

LACGUS

A P

M l

M 2

T R

P I

P2

Total

LAC GUS-

A P

M l

M2

TR

PI

P2

Total

Isolates

N o

37

31

42

41

31

11

193

24

29

48

48

52

11

212

2

9

0

3

2

0

16

17

23

15

36

46

29

166

E

N o

14

10

25

19

18

1

87

24

27

48

46

52

10

207

2

8

0

3

2

0

15

0

2

1

5

1

0

9

coli

( )

38

32

59

46

58

9

45

100

93

100

96

100

91

98

100

89

0

100

100

0

94

0

9

7

14

2

0

5

K E C

No

23

20

17

17

8

1

86

0

2

0

2

0

1

5

0

1

0

0

0

0

1

17

20

12

14

6

2

71

( )

62

68

41

42

26

9

45

0

7

0

4

0

9

2

0

11

0

0

0

0

6

100

87

80

39

13

7

43

Non

coliform

b

No

0

0

0

0

3

5

8

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

1

2

12

22

10

47

( )

0

0

0

0

10

46

4

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

4

13

33

48

34

28

N ot

identified

No

0

1

0

5

2

4

12

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

5

17

17

39

( )

0

3

0

12

6

36

6

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

14

37

59

24

KEC

Klebsiella Enterobacter and Citrobacter

b

Oxidase + P seudomonas spp Vibrio spp Aeromonas spp

Sampling sites



i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5



TAB LE 5

Identif ication o f Colonies Picked from C C Agar Using the A PI 20E System

O

m

sect

o

L A C

+

G U S

N o

L A C

+

G U S N o L A C G U S

4

N o L A C G U S -

No


Klebsiella oxytoca

K Pneuntoniae

Citrobacterfreundii

C Amalonaticus

Escherichia coli

4

2

2

6

1

6

pound co

Cfreundii

8

1

ot l

E co

21

Pseudomonas

spp

P fluorescens

Acinetobacter

spp

Flavobacterium

spp

Proteus

spp

Salmonella typhi

Citrobacterfreundii

C amalonaticus

Klebsiella oxytoca

K pneumoniae


E agglomerans

E sakazakii

Escherichia coli

4

1

1

1

1

1

8

1

4

2

3

1

1

4

33

a

LA C

+

G US salmon to red colonies

bull LAC GU S

+

dark-blue to violet colonies

l A C

G US

+

light-blue to turquoise colonies

d

L AC GU S colourless colonies D o

w n l o a d e d b y [ S o u t h e r n I l l i n o i s U n i v e r s i t y ] a t 2 1 0 5 1 8 J u l y 2 0 1 5



1242 ALONSO ET AL

to transport the glucuronide substrate across the cell membrane (Coyne and Schuler

1994) Som e authors (Bej et al 1 991 Cleuziat and Rob ert-Baud oy 1990 Fen g et al

1991

Flicker and Flicker 1994 Green et al 19 91 M artins et al 199 3

Venkateswaran et al 1996) observed that part of the genetic sequences of the uidA

gene which encodes for the GUS enzyme was present in most if not all E coli

isolates regardless of the GUS phenotype Frampton and Restaino (1993) indicated

that the following factors may influence the GUS assay substantially whichever GUS

detection system is used strain differences in response to particular substrates and

substrate concentration effects of carbohydrate content and selective agents in the

medium incubation time and temperature pH changes ionic strength effects and

possible interference by large numbers of competing bacteria or substances in the

sample

itself

W e have isolated one strain of Citrobacter freundii LAC

+

G U S

+

Although P-glucuronidase activity has been reported in some strains of coliforms

(Enterobacter agg lomerans E cloacae E amnig enus C itrobacter freundii C

amalonaticus Escherichia vulneris and Hqfnia alvet) Aeromonas sp and

Acinetobacter sp (Heizmann 1988 Kaacutempfer et al 199 1 Perez et al 1986 Sartory y

Howard 1992 Watkins et al 1988) their occurrence appears to be very infrequent

(Sartory and Howard 1992) The reason for the production of p-glucuronidase by

these strains is not know n but o ther investigators (Brenner et al 1993) have sugge sted

tha t the reaction ma y be plasmid mediated

The specificity of the medium for KEC coliforms was low Of the 193 salmon to

red colonies (LAC

+

GUS ) 86 (45) were confirmed as KEC coliforms giving a false

positive rate of 5 5 (127 of 193 colonies) A total of

7

LAC GUS colonies 1 LAC


i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5




G U S

+

colony and 5 LAC

+

GUS

+

colonies were KEC coliforms resulting in a false

negative rate of 195 (77 of 394 colonies) A high incidence of false negative (L A C )

KEC colonies was detected Because enzyme activities are subject to the physiological

status o f the bacteria a variable fraction of the coliform bacteria m ay be stressed wh en

changes in irradiation salinity temperature and nutrient concentration of the

environment occur (Pommepuy et al 1992) Fecal coliform bacteria comprise several

bacterial species and their response to environmental factors may not be the same for

each species (Pommepuy et al 1996) In treated drinking water injured coliforms can

comprise between 50 and gt90 of coliforms present (McFeters 1989) A total of 31

L A C G U S colonies were streaked onto CC agar and incubated at 37degC 29 K EC

strains that failed to produce P-galactosidase at 445degC were able to produce the

enzyme at 37degC Dockins and McFeters (1978) observed that optimal activity of 0-

galactosidase enzyme in freshly sonic extracts fecal coliforms typically occurred at

30+2degC and the activity decreased rapidly as the temperature increased above 35 to

38degC At 445degC fecal P-galactosidase activity was 25 to 50 of the optimal

tem peratu re (D ockins and M cFeters 1978) This decrease in p-galactosidase activity in

fecal coliforms has been indirectly observed by Warren et al (Warren et al 1976) who

found that lowering the 445degC incubation temperature by 1 or 2degC resulted in

significantly faster rate of ONPG hydrolysis Munro et al (1987) observed that P-

galactosidase activity of pound coli starved cells disappeared gradually with time The

physiological condition of KEC isolates could be responsible for the non-expression of

enzym e activity at 445degC


i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5



1244 ALONSO ET AL

When LAC

+

GUS LAC

+

GUS

+

and LAC GUS colonies were considered as

fecal coliforms (included E coli) more than 95 (401 of 421 colonies) of the

identified colonies belonged to the fecal coliform group giving a false positive rate of

4 8 (20 of 421 colonies) Nevertheless LAC GUS colonies represented 481 (80

of 166 colonies) of the identified coliform gro up

Results of the study indicated that 94 (205 of 219 colonies) of the E coli

LAC

+

GUS strains produced gas in the EC medium (Table 6) Thermotolerant E coli

was the most frequently isolated in the 6 environmental conditions as expected

Ho we ver the percentage was variably ranging from 8 2 (P2) to 100 (AP ) A total

of 219 E coli strains (LAC

+

GUS) were verified in EC broth and 12 (5) gas

negative strains were encountered In EC broth K coli must transport lactose throug h

the cell membrane transform the substrate to glucose metabolize glucose through the

glycolytic cycle to pyruvate and then convert pyruvate to the desired end prod uct

either acid or gas (Edberg et al 1988) Because lactose fermentation at 445degC is

determined by a complex of different enzymes a number of anomalous results may

occur such as false negative gas production (Edberg et al 1988 Gtammanco et al

1992)

Leclerc et al (1977) observed that the activity of formic h ydrogen lyase which

is needed for gas production from lactose is quite often reduced and sometimes

entirely suppressed under conditions that do not favour survival of coliforms in water

Munro et al (1987) suggested that the disappearance of P-galactosidase activity in

non-salt adapted

E coli

cells starved in seawater could have implications for their

enumeration by standard cultural methods all of which being grounded on the

acidification and fermentation of lactose






TABLEOacute

Percentage of Thermotolerant ThermosensUive and iacutendo le Negative

K coli

LAC GUS) Strains Recovered in C C A ga r

Sampling

sites

AP

M l

M2

TR

PI

P2

N o of

strains

24

30

53

46

55

11

Thermo-

tolerant

No

24

28

51

42

51

9

100

93

96

91

93

82

Thermo-

sensitive

b

No

0

2

2

4

3

12

0

7

4

9

5

5

Indol-

No

0

2

3

2

1

9

0

7

6

4

2

4

Therm otolerant gas formed from lactose a 1445degC

k

Therm osensitive gas not formed from lactose at 445degC

The data obtained suggested that specificity of CC agar for fecal coliforms was

related to the incubation temperature and we are of the opinion that lowering the

445 degC incubation temp erature to 41degC may reverse the expression of P-galactosidase

and P-glucuron idase activities of som e metabolically injured fecal co liforms

REFERENCES

Alonso JL Amoros I Chong S and Garelick H J Microbiol Methods 25 309-

315(1996)

AP HA Standard Metho ds for the Examination of W ater and W astewater 19th

edition Am erican Public Health Association N ew Y ork (1995 ) 9 pp

1-117

Augo ustinos MT Grabow NA and Kfir R W at Sci Tech 27 267-270 (1993)

Bej

AK McCarty SC and Atlas RM Appl Environ Microbiol 57 2429-2432

(1991)


i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5



1246 ALONSO ET AL

Brenner KP Rankin CC Roybal YR and Stelma JrG Appl Environ

M icrobiol 59 3534-3544 (1993)

Brodsky MH A SM N ews 63 345-346 (1997)

Caplenas NR and Kanarek MS Am J Publ Hlth 74 1273-1275 (1984)

Charriere G Mossel DAA Beaudeau P and Leclerc H J Appl Bacteriol 76

336-344 (1992)

Ciebin BW Brod sky MH Edding ton R Horsnell G Choney A Palmateer G

Ley A Joshi R and Shears G Appl Environ M icrobiol 6 1 3940-3942 (1995)

Clark DL Milner BB Stewart MH Wolfe RL and Olson BH Appl Environ

Microbiol 57 1528-1534(1991)

Cleuziat P and Robert-Baudoy J FEMS Microbiol Lett 72 315-322 (1990)

Cov ert TC Rice EW Johnson SA B erman D Johnson CH and Mason PJ

J A W W A 84 98-104 (1992)

Coy ne M S and Schuler J C J Environ Qu al 2 3 126-129 (1994 )

Do ckins W S and M cFeters GA Appl Environ Microbiol 36 341-348 (1978 )

Dufour A P Bacterial IndicatorsHealth Haz ards Associated with W ater Ed AW

Ho adley and BJ Dutka American Society for Testmg M aterials Philadelphia (197 7)

pp 48-58

Edberg SC Allen M J Smith D B and the National Collaborative Study Appl

Environ Microbiol 54 1595-1601 (1988)

Evison LM Wat Sci Tech 20 309-315 (1988)

Feng P Lum R and Chang G W Appl Environ M icrobiol 5 7 320-323 (1991)

Figueras M J Po lo F Inza I and Guarro J Lett Appl M icrobiol 19 446-450

(1994)

Frampton EW and Restaino L J Appl Ba cterio l 74 223 -233

Fricker EJ and Fricker CR Lett Appl M icrobiol 19 44-46 (199 4)

Giammanco G Pignato S and Biondi M Zbl H y g 193 99-105 (1992)



i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5



QUANTITATIVE DETERMINATION OF

E COLI

1247

Green DH Lewis GD Rodtong S and Loutit MW J Microbiol Methods 13

207-214 (1991)

Heizmann W Doumlller P C Guumltbrod B and W erner H J Clin Microbiol 26 268 2-

2684 (1988)

Kampfer P

Rauhoff

D and D ott W J Clin M icrobiol 29 2877-28 79 (1 991 )

Lec lerc H M ossel D A A Trinel PA and Gavini F Bacterial IndicatorsHealth

Haza rd A ssociated with Water Ed AW Hoadley and BJ Dutka Am erican Society

for Testing M aterials Philadelphia (1977) pp 22 -36

Manafi M Kneifel W and Bascomb S Microbiol R e v 55 335-348 (1991)

M artins M T Rivera IG Clark D L Stewart M H W olfe RL and Olsen B H

Appl Environ M icrob iol 5 9 2271-2276 (199 3)

M cFe ters G A Injured index and pathogenic bacteria occurrence and detection in

foods

w ater and feeds Ed B Ray CRC Press Bo ca Raton (1989) pp 179-210

Mossel D A A A SM New s 63 175 (1997)

M un ro P M Gauthier MJ and Laumond F M Appl Environ Micro biol 53

1476-1481 (1987)

Palmer C J Tsai Y L ang AL and Sangermano L R A ppl Environ M icrob iol

59 786-790(1995)

Perez J L Berrocal CI and Berrocal L J Appl Ba cteriol 6 1 541-545 (19 86)

Pom mepuy M Guillard J F Duprey E D errien A Le Guyader F and Cormier

M W at Sci T ec h 2 5 93-103 (1992)

Pomm epuy M Fiksdal L Gourmelon M M elikechi H Caprais M P Cormier

M and Colwell RR J Appl Ba cterio l 8 1 174-180 (1996)

Pressw ood W G and Strong D K A ppl Environ M icrobiol 36 90-94 (1978 )

Rice

E W Allen MJ and Edberg SC Appl Environ M icrob iol 56 1203-1205

(1990)

Ro se R E Geldreich EE and Litsky W Appl M icrobiol 29 532-536 (1975)

Sartory DP and How ard L Lett Appl M icrobiol 15 273-276 (1992)





1248 ALONSO ET AL

Shadix LC Dunnigan M E and Rice E W Can J M icrobiol 39 1066-1070

(1993)

Venkateswaran K Murakoshi A and Satake M Appl Environ Microbiol 62

2236-2243 (1996)

Warren LS Benoit RE and Jessee JA Appl Environ Microbiol 35 136-141

(1976)

Watkins WD Rippey SR Clavet CR Kelley-Reitz D J and Burkhardt W

Appl Environ Microbiol 54 1874-1875 (1988)

e c e i v e d

December

22 1997


i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5



PLEASE SCROLL DOWN FOR ARTICLE

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licensors make no representations or warranties whatsoever as to theaccuracy completeness or suitability for any purpose of the ContentAny opinions and views expressed in this publication are the opinionsand views of the authors and are not the views of or endorsed byTaylor amp Francis The accuracy of the Content should not be reliedupon and should be independently verified with primary sources of information Taylor and Francis shall not be liable for any lossesactions claims proceedings demands costs expenses damages

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and-conditions

D o w n l o a d e d b y [

S o u t h e r n I l l i n o i s U n i v e r s i t y ] a t 2 1 0 5 1 8 J u l y 2 0 1

5





COLIFORMS IN

W TER

USING CHROMOGENIC

M E I U M




J L Alonso

1

A Soriano

2

I

Amoros

1

and MA

F e r rus

3

1

Inst i tu to


46022 Valencia

2


3


Spain

STR CT








β 983085

1229



i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5



1230 ALONSO ET AL

glucuronidase



incubation



coli



false


E

coli (296 )

colonies


K

coli



G U S

+


streaked on to

CC





INTRODUCTION

The


importance



The


tha t






Citrobacter)

may














E coli





(APHA 1995)






+

GUS ) and the




+

GUS)






i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5



1232 ALONSO ET AL


Sampling












K coli




Bacterial Strains














TABLE 1



1233

Test Strain






Klebsiella oxytoca

Klebsiella ozaenae




Escherichia coli

Hafnia alvei


Serratia marcescens

Serratia rubiadea

Cedecea davisae

Kluyvera ascorbata

Shigella flexneri

Shigella boydii

Shigella sonnei

Aeromonas hydrofila

Aeromonas caviae

Aeromonas media

A eromonas jandaei


Aeromonas trota


A

veronii bv veronii

Vibrio cholerae



Enterococcusfaecium

Salmonella derby


Salmonella london

No

684

194

858

857

140

860

851

856

863

401

678

157

867

159

868

842

861

585

583

413

398

838

4232

4228

4241

4255

4224

4257

557

108

184

410

IHMN

IHMN

IHMN

37degC

G

b

C

c

+ r

+ r

+ r

+ r

+ r

+ r

+ r

+ r

+ r

+ r

+ b

+ r

+ r

+ r

+ r

+ c

+ r

+ c

+ t

+ b

+ r

+ r

+ r

+ r

+ c

+ r

+ r

+ r

+ r

+ c

-

-

+ c

+ t

+ t

41

G ~

+

+

+

+

+

+

+

+

+

+

+

+

+

+

-

+

+

+

+

+

+

+

+

-

+

+

+

+

+

-

-

+

+

+

degC

r

r

r

r

r

r

r

r

c

r

b

r

r

t

r

-

r

c

t

b

r

r

r

r

-

r

r

r

r

c

-

-

c

t

t

445degC

G C

+ r

+ r

+ r

-

+ c

+ r

+ r

+ c

+ c

+ r

+ b

+ r

+ r

+ r

+ r

-

+ r

+ c

+ t

+ b

-

-

+ r

-

-

-

-

-

+ c

-

-

+ c

+ t

+ t


G


b

C


c=Colourless





1234 ALONSO ET AL












+



K coli

For














i t y ] a t 2 1 0 5 1 8 J u l y 2 0 1 5






(LAC

+


4








system (bioMerieux)















i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5



1236 ALONSO ET AL

TABLE 2

N o n - pound

coli

Fecal Coliforms

Klebsiella

spp

Enterobacter

spp and

Citrobacter

spp) KE C ) and

Escherich ia coli



Sampling

source

TR

EC-CCA

b

KEC-CCA

C

EKEC-CCA

d

F C - m F C

PI

EC-CCA

KEC-CCA

EKEC-CCA

FC-mFC

AP

E C - C C A

K E C - C C A

EKEC-CCA

FC-mFC

M l

EC-CCA

K E C - C C A

EKEC-CCA

FC-mFC

M2

EC-CCA

K E C - C C A

E K E C - C C A

FC-mFC

Mean

215

197

239

245

195

081

150

168

672

596

679

673

535

451

541

536

307

253

318

313

2h37deg-445degC

SD

151

163

156

167

062

025

080

071

020

014

019

019

124

112

122

125

093

083

091

098

Min

070

030

085

085

111

048

048

060

646

578

656

652

338

270

346

336

208

154

220

208

Max

408

420

445

461

258

108

259

262

700

623

705

703

672

570

676

671

448

370

454

462

Mean

287

264

237

241

179

060

161

161

671

582

676

673

532

439

537

536

300

240

311

307

445

SD

141

148

157

158

055

026

072

077

019

016

018

019

122

129

123

123

102

085

098

101

degC

Min

158

100

078

104

118

030

048

048

651

560

657

652

338

230

341

338

194

140

205

181

Max

426

420

453

448

250

095

250

256

699

611

703

704

669

604

678

668

457

378

463

458



and maximum (Max)



+


d




i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5







s
























1238

ALONSO ET AL

T A B L E 3



Sample

site

T R

P I

AP

M l

M 2

Parameters

EC37-KEC37

EC44-KEC44

EC37-KEC37

EC44-KEC44

EC37-KEC37

EC44-KEC44

EC37-KEC37

EC44-KEC44

EC37-KEC37

EC44-KEC44

R

099

098

069

064

068

040

099

099

099

098

P

lt001

lt001

N S

b

N S

N S

N S

lt001

lt001

lt001

lt001

Intercept

(a)

0340

0405

0571

1729

0977

3981

0419

1185

0257

0170

Slope

(b )

0916

0936

1701

0108

0963

0468

1093

0941

1112

0182

a












Aeromonas jandaei


i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5







+

G U S L A C

+

G U S

+

LAC



+

GUD species




+

GUS

4



15 L A C G U S

+


+


















i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5



24

ALONSO ET AL

TABLE 4

Num ber o f

coli

a nd N o n-E

coli



Reactions

Phenotype

LAC GUS

AP

C

M l

M 2

TR

PI

P2

Total

LAC

+

GUS

+

AP

M l

M 2

T R

PI

P2

Total

LACGUS

A P

M l

M 2

T R

P I

P2

Total

LAC GUS-

A P

M l

M2

TR

PI

P2

Total

Isolates

N o

37

31

42

41

31

11

193

24

29

48

48

52

11

212

2

9

0

3

2

0

16

17

23

15

36

46

29

166

E

N o

14

10

25

19

18

1

87

24

27

48

46

52

10

207

2

8

0

3

2

0

15

0

2

1

5

1

0

9

coli

( )

38

32

59

46

58

9

45

100

93

100

96

100

91

98

100

89

0

100

100

0

94

0

9

7

14

2

0

5

K E C

No

23

20

17

17

8

1

86

0

2

0

2

0

1

5

0

1

0

0

0

0

1

17

20

12

14

6

2

71

( )

62

68

41

42

26

9

45

0

7

0

4

0

9

2

0

11

0

0

0

0

6

100

87

80

39

13

7

43

Non

coliform

b

No

0

0

0

0

3

5

8

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

1

2

12

22

10

47

( )

0

0

0

0

10

46

4

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

4

13

33

48

34

28

N ot

identified

No

0

1

0

5

2

4

12

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

5

17

17

39

( )

0

3

0

12

6

36

6

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

14

37

59

24

KEC


b


Sampling sites



i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5



TAB LE 5


O

m

sect

o

L A C

+

G U S

N o

L A C

+


4

N o L A C G U S -

No


Klebsiella oxytoca

K Pneuntoniae

Citrobacterfreundii

C Amalonaticus

Escherichia coli

4

2

2

6

1

6

pound co

Cfreundii

8

1

ot l

E co

21

Pseudomonas

spp

P fluorescens

Acinetobacter

spp

Flavobacterium

spp

Proteus

spp

Salmonella typhi

Citrobacterfreundii

C amalonaticus

Klebsiella oxytoca

K pneumoniae


E agglomerans

E sakazakii

Escherichia coli

4

1

1

1

1

1

8

1

4

2

3

1

1

4

33

a

LA C

+


bull LAC GU S

+


l A C

G US

+


d





1242 ALONSO ET AL



1991










sample

itself


+

G U S

+










red colonies (LAC

+



7



i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5




G U S

+

colony and 5 LAC

+

GUS

+
























i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5



1244 ALONSO ET AL

When LAC

+

GUS LAC

+

GUS

+







LAC

+





+








1992)





non-salt adapted

E coli









TABLEOacute


K coli


Sampling

sites

AP

M l

M2

TR

PI

P2

N o of

strains

24

30

53

46

55

11

Thermo-

tolerant

No

24

28

51

42

51

9

100

93

96

91

93

82

Thermo-

sensitive

b

No

0

2

2

4

3

12

0

7

4

9

5

5

Indol-

No

0

2

3

2

1

9

0

7

6

4

2

4


k






REFERENCES


315(1996)



1-117


Bej


(1991)


i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5



1246 ALONSO ET AL


M icrobiol 59 3534-3544 (1993)




336-344 (1992)




Microbiol 57 1528-1534(1991)



J A W W A 84 98-104 (1992)





pp 48-58






(1994)






i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5




E COLI

1247


207-214 (1991)


2684 (1988)

Kampfer P

Rauhoff









foods




1476-1481 (1987)


59 786-790(1995)



M W at Sci T ec h 2 5 93-103 (1992)




Rice


(1990)







1248 ALONSO ET AL


(1993)


2236-2243 (1996)


(1976)



e c e i v e d

December

22 1997


i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5





COLIFORMS IN

W TER

USING CHROMOGENIC

M E I U M




J L Alonso

1

A Soriano

2

I

Amoros

1

and MA

F e r rus

3

1

Inst i tu to


46022 Valencia

2


3


Spain

STR CT








β 983085

1229



i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5



1230 ALONSO ET AL

glucuronidase



incubation



coli



false


E

coli (296 )

colonies


K

coli



G U S

+


streaked on to

CC





INTRODUCTION

The


importance



The


tha t






Citrobacter)

may














E coli





(APHA 1995)






+

GUS ) and the




+

GUS)






i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5



1232 ALONSO ET AL


Sampling












K coli




Bacterial Strains














TABLE 1



1233

Test Strain






Klebsiella oxytoca

Klebsiella ozaenae




Escherichia coli

Hafnia alvei


Serratia marcescens

Serratia rubiadea

Cedecea davisae

Kluyvera ascorbata

Shigella flexneri

Shigella boydii

Shigella sonnei

Aeromonas hydrofila

Aeromonas caviae

Aeromonas media

A eromonas jandaei


Aeromonas trota


A

veronii bv veronii

Vibrio cholerae



Enterococcusfaecium

Salmonella derby


Salmonella london

No

684

194

858

857

140

860

851

856

863

401

678

157

867

159

868

842

861

585

583

413

398

838

4232

4228

4241

4255

4224

4257

557

108

184

410

IHMN

IHMN

IHMN

37degC

G

b

C

c

+ r

+ r

+ r

+ r

+ r

+ r

+ r

+ r

+ r

+ r

+ b

+ r

+ r

+ r

+ r

+ c

+ r

+ c

+ t

+ b

+ r

+ r

+ r

+ r

+ c

+ r

+ r

+ r

+ r

+ c

-

-

+ c

+ t

+ t

41

G ~

+

+

+

+

+

+

+

+

+

+

+

+

+

+

-

+

+

+

+

+

+

+

+

-

+

+

+

+

+

-

-

+

+

+

degC

r

r

r

r

r

r

r

r

c

r

b

r

r

t

r

-

r

c

t

b

r

r

r

r

-

r

r

r

r

c

-

-

c

t

t

445degC

G C

+ r

+ r

+ r

-

+ c

+ r

+ r

+ c

+ c

+ r

+ b

+ r

+ r

+ r

+ r

-

+ r

+ c

+ t

+ b

-

-

+ r

-

-

-

-

-

+ c

-

-

+ c

+ t

+ t


G


b

C


c=Colourless





1234 ALONSO ET AL












+



K coli

For














i t y ] a t 2 1 0 5 1 8 J u l y 2 0 1 5






(LAC

+


4








system (bioMerieux)















i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5



1236 ALONSO ET AL

TABLE 2

N o n - pound

coli

Fecal Coliforms

Klebsiella

spp

Enterobacter

spp and

Citrobacter

spp) KE C ) and

Escherich ia coli



Sampling

source

TR

EC-CCA

b

KEC-CCA

C

EKEC-CCA

d

F C - m F C

PI

EC-CCA

KEC-CCA

EKEC-CCA

FC-mFC

AP

E C - C C A

K E C - C C A

EKEC-CCA

FC-mFC

M l

EC-CCA

K E C - C C A

EKEC-CCA

FC-mFC

M2

EC-CCA

K E C - C C A

E K E C - C C A

FC-mFC

Mean

215

197

239

245

195

081

150

168

672

596

679

673

535

451

541

536

307

253

318

313

2h37deg-445degC

SD

151

163

156

167

062

025

080

071

020

014

019

019

124

112

122

125

093

083

091

098

Min

070

030

085

085

111

048

048

060

646

578

656

652

338

270

346

336

208

154

220

208

Max

408

420

445

461

258

108

259

262

700

623

705

703

672

570

676

671

448

370

454

462

Mean

287

264

237

241

179

060

161

161

671

582

676

673

532

439

537

536

300

240

311

307

445

SD

141

148

157

158

055

026

072

077

019

016

018

019

122

129

123

123

102

085

098

101

degC

Min

158

100

078

104

118

030

048

048

651

560

657

652

338

230

341

338

194

140

205

181

Max

426

420

453

448

250

095

250

256

699

611

703

704

669

604

678

668

457

378

463

458



and maximum (Max)



+


d




i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5







s
























1238

ALONSO ET AL

T A B L E 3



Sample

site

T R

P I

AP

M l

M 2

Parameters

EC37-KEC37

EC44-KEC44

EC37-KEC37

EC44-KEC44

EC37-KEC37

EC44-KEC44

EC37-KEC37

EC44-KEC44

EC37-KEC37

EC44-KEC44

R

099

098

069

064

068

040

099

099

099

098

P

lt001

lt001

N S

b

N S

N S

N S

lt001

lt001

lt001

lt001

Intercept

(a)

0340

0405

0571

1729

0977

3981

0419

1185

0257

0170

Slope

(b )

0916

0936

1701

0108

0963

0468

1093

0941

1112

0182

a












Aeromonas jandaei


i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5







+

G U S L A C

+

G U S

+

LAC



+

GUD species




+

GUS

4



15 L A C G U S

+


+


















i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5



24

ALONSO ET AL

TABLE 4

Num ber o f

coli

a nd N o n-E

coli



Reactions

Phenotype

LAC GUS

AP

C

M l

M 2

TR

PI

P2

Total

LAC

+

GUS

+

AP

M l

M 2

T R

PI

P2

Total

LACGUS

A P

M l

M 2

T R

P I

P2

Total

LAC GUS-

A P

M l

M2

TR

PI

P2

Total

Isolates

N o

37

31

42

41

31

11

193

24

29

48

48

52

11

212

2

9

0

3

2

0

16

17

23

15

36

46

29

166

E

N o

14

10

25

19

18

1

87

24

27

48

46

52

10

207

2

8

0

3

2

0

15

0

2

1

5

1

0

9

coli

( )

38

32

59

46

58

9

45

100

93

100

96

100

91

98

100

89

0

100

100

0

94

0

9

7

14

2

0

5

K E C

No

23

20

17

17

8

1

86

0

2

0

2

0

1

5

0

1

0

0

0

0

1

17

20

12

14

6

2

71

( )

62

68

41

42

26

9

45

0

7

0

4

0

9

2

0

11

0

0

0

0

6

100

87

80

39

13

7

43

Non

coliform

b

No

0

0

0

0

3

5

8

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

1

2

12

22

10

47

( )

0

0

0

0

10

46

4

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

4

13

33

48

34

28

N ot

identified

No

0

1

0

5

2

4

12

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

5

17

17

39

( )

0

3

0

12

6

36

6

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

14

37

59

24

KEC


b


Sampling sites



i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5



TAB LE 5


O

m

sect

o

L A C

+

G U S

N o

L A C

+


4

N o L A C G U S -

No


Klebsiella oxytoca

K Pneuntoniae

Citrobacterfreundii

C Amalonaticus

Escherichia coli

4

2

2

6

1

6

pound co

Cfreundii

8

1

ot l

E co

21

Pseudomonas

spp

P fluorescens

Acinetobacter

spp

Flavobacterium

spp

Proteus

spp

Salmonella typhi

Citrobacterfreundii

C amalonaticus

Klebsiella oxytoca

K pneumoniae


E agglomerans

E sakazakii

Escherichia coli

4

1

1

1

1

1

8

1

4

2

3

1

1

4

33

a

LA C

+


bull LAC GU S

+


l A C

G US

+


d





1242 ALONSO ET AL



1991










sample

itself


+

G U S

+










red colonies (LAC

+



7



i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5




G U S

+

colony and 5 LAC

+

GUS

+
























i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5



1244 ALONSO ET AL

When LAC

+

GUS LAC

+

GUS

+







LAC

+





+








1992)





non-salt adapted

E coli









TABLEOacute


K coli


Sampling

sites

AP

M l

M2

TR

PI

P2

N o of

strains

24

30

53

46

55

11

Thermo-

tolerant

No

24

28

51

42

51

9

100

93

96

91

93

82

Thermo-

sensitive

b

No

0

2

2

4

3

12

0

7

4

9

5

5

Indol-

No

0

2

3

2

1

9

0

7

6

4

2

4


k






REFERENCES


315(1996)



1-117


Bej


(1991)


i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5



1246 ALONSO ET AL


M icrobiol 59 3534-3544 (1993)




336-344 (1992)




Microbiol 57 1528-1534(1991)



J A W W A 84 98-104 (1992)





pp 48-58






(1994)






i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5




E COLI

1247


207-214 (1991)


2684 (1988)

Kampfer P

Rauhoff









foods




1476-1481 (1987)


59 786-790(1995)



M W at Sci T ec h 2 5 93-103 (1992)




Rice


(1990)







1248 ALONSO ET AL


(1993)


2236-2243 (1996)


(1976)



e c e i v e d

December

22 1997


i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5



1230 ALONSO ET AL

glucuronidase



incubation



coli



false


E

coli (296 )

colonies


K

coli



G U S

+


streaked on to

CC





INTRODUCTION

The


importance



The


tha t






Citrobacter)

may














E coli





(APHA 1995)






+

GUS ) and the




+

GUS)






i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5



1232 ALONSO ET AL


Sampling












K coli




Bacterial Strains














TABLE 1



1233

Test Strain






Klebsiella oxytoca

Klebsiella ozaenae




Escherichia coli

Hafnia alvei


Serratia marcescens

Serratia rubiadea

Cedecea davisae

Kluyvera ascorbata

Shigella flexneri

Shigella boydii

Shigella sonnei

Aeromonas hydrofila

Aeromonas caviae

Aeromonas media

A eromonas jandaei


Aeromonas trota


A

veronii bv veronii

Vibrio cholerae



Enterococcusfaecium

Salmonella derby


Salmonella london

No

684

194

858

857

140

860

851

856

863

401

678

157

867

159

868

842

861

585

583

413

398

838

4232

4228

4241

4255

4224

4257

557

108

184

410

IHMN

IHMN

IHMN

37degC

G

b

C

c

+ r

+ r

+ r

+ r

+ r

+ r

+ r

+ r

+ r

+ r

+ b

+ r

+ r

+ r

+ r

+ c

+ r

+ c

+ t

+ b

+ r

+ r

+ r

+ r

+ c

+ r

+ r

+ r

+ r

+ c

-

-

+ c

+ t

+ t

41

G ~

+

+

+

+

+

+

+

+

+

+

+

+

+

+

-

+

+

+

+

+

+

+

+

-

+

+

+

+

+

-

-

+

+

+

degC

r

r

r

r

r

r

r

r

c

r

b

r

r

t

r

-

r

c

t

b

r

r

r

r

-

r

r

r

r

c

-

-

c

t

t

445degC

G C

+ r

+ r

+ r

-

+ c

+ r

+ r

+ c

+ c

+ r

+ b

+ r

+ r

+ r

+ r

-

+ r

+ c

+ t

+ b

-

-

+ r

-

-

-

-

-

+ c

-

-

+ c

+ t

+ t


G


b

C


c=Colourless





1234 ALONSO ET AL












+



K coli

For














i t y ] a t 2 1 0 5 1 8 J u l y 2 0 1 5






(LAC

+


4








system (bioMerieux)















i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5



1236 ALONSO ET AL

TABLE 2

N o n - pound

coli

Fecal Coliforms

Klebsiella

spp

Enterobacter

spp and

Citrobacter

spp) KE C ) and

Escherich ia coli



Sampling

source

TR

EC-CCA

b

KEC-CCA

C

EKEC-CCA

d

F C - m F C

PI

EC-CCA

KEC-CCA

EKEC-CCA

FC-mFC

AP

E C - C C A

K E C - C C A

EKEC-CCA

FC-mFC

M l

EC-CCA

K E C - C C A

EKEC-CCA

FC-mFC

M2

EC-CCA

K E C - C C A

E K E C - C C A

FC-mFC

Mean

215

197

239

245

195

081

150

168

672

596

679

673

535

451

541

536

307

253

318

313

2h37deg-445degC

SD

151

163

156

167

062

025

080

071

020

014

019

019

124

112

122

125

093

083

091

098

Min

070

030

085

085

111

048

048

060

646

578

656

652

338

270

346

336

208

154

220

208

Max

408

420

445

461

258

108

259

262

700

623

705

703

672

570

676

671

448

370

454

462

Mean

287

264

237

241

179

060

161

161

671

582

676

673

532

439

537

536

300

240

311

307

445

SD

141

148

157

158

055

026

072

077

019

016

018

019

122

129

123

123

102

085

098

101

degC

Min

158

100

078

104

118

030

048

048

651

560

657

652

338

230

341

338

194

140

205

181

Max

426

420

453

448

250

095

250

256

699

611

703

704

669

604

678

668

457

378

463

458



and maximum (Max)



+


d




i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5







s
























1238

ALONSO ET AL

T A B L E 3



Sample

site

T R

P I

AP

M l

M 2

Parameters

EC37-KEC37

EC44-KEC44

EC37-KEC37

EC44-KEC44

EC37-KEC37

EC44-KEC44

EC37-KEC37

EC44-KEC44

EC37-KEC37

EC44-KEC44

R

099

098

069

064

068

040

099

099

099

098

P

lt001

lt001

N S

b

N S

N S

N S

lt001

lt001

lt001

lt001

Intercept

(a)

0340

0405

0571

1729

0977

3981

0419

1185

0257

0170

Slope

(b )

0916

0936

1701

0108

0963

0468

1093

0941

1112

0182

a












Aeromonas jandaei


i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5







+

G U S L A C

+

G U S

+

LAC



+

GUD species




+

GUS

4



15 L A C G U S

+


+


















i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5



24

ALONSO ET AL

TABLE 4

Num ber o f

coli

a nd N o n-E

coli



Reactions

Phenotype

LAC GUS

AP

C

M l

M 2

TR

PI

P2

Total

LAC

+

GUS

+

AP

M l

M 2

T R

PI

P2

Total

LACGUS

A P

M l

M 2

T R

P I

P2

Total

LAC GUS-

A P

M l

M2

TR

PI

P2

Total

Isolates

N o

37

31

42

41

31

11

193

24

29

48

48

52

11

212

2

9

0

3

2

0

16

17

23

15

36

46

29

166

E

N o

14

10

25

19

18

1

87

24

27

48

46

52

10

207

2

8

0

3

2

0

15

0

2

1

5

1

0

9

coli

( )

38

32

59

46

58

9

45

100

93

100

96

100

91

98

100

89

0

100

100

0

94

0

9

7

14

2

0

5

K E C

No

23

20

17

17

8

1

86

0

2

0

2

0

1

5

0

1

0

0

0

0

1

17

20

12

14

6

2

71

( )

62

68

41

42

26

9

45

0

7

0

4

0

9

2

0

11

0

0

0

0

6

100

87

80

39

13

7

43

Non

coliform

b

No

0

0

0

0

3

5

8

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

1

2

12

22

10

47

( )

0

0

0

0

10

46

4

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

4

13

33

48

34

28

N ot

identified

No

0

1

0

5

2

4

12

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

5

17

17

39

( )

0

3

0

12

6

36

6

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

14

37

59

24

KEC


b


Sampling sites



i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5



TAB LE 5


O

m

sect

o

L A C

+

G U S

N o

L A C

+


4

N o L A C G U S -

No


Klebsiella oxytoca

K Pneuntoniae

Citrobacterfreundii

C Amalonaticus

Escherichia coli

4

2

2

6

1

6

pound co

Cfreundii

8

1

ot l

E co

21

Pseudomonas

spp

P fluorescens

Acinetobacter

spp

Flavobacterium

spp

Proteus

spp

Salmonella typhi

Citrobacterfreundii

C amalonaticus

Klebsiella oxytoca

K pneumoniae


E agglomerans

E sakazakii

Escherichia coli

4

1

1

1

1

1

8

1

4

2

3

1

1

4

33

a

LA C

+


bull LAC GU S

+


l A C

G US

+


d





1242 ALONSO ET AL



1991










sample

itself


+

G U S

+










red colonies (LAC

+



7



i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5




G U S

+

colony and 5 LAC

+

GUS

+
























i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5



1244 ALONSO ET AL

When LAC

+

GUS LAC

+

GUS

+







LAC

+





+








1992)





non-salt adapted

E coli









TABLEOacute


K coli


Sampling

sites

AP

M l

M2

TR

PI

P2

N o of

strains

24

30

53

46

55

11

Thermo-

tolerant

No

24

28

51

42

51

9

100

93

96

91

93

82

Thermo-

sensitive

b

No

0

2

2

4

3

12

0

7

4

9

5

5

Indol-

No

0

2

3

2

1

9

0

7

6

4

2

4


k






REFERENCES


315(1996)



1-117


Bej


(1991)


i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5



1246 ALONSO ET AL


M icrobiol 59 3534-3544 (1993)




336-344 (1992)




Microbiol 57 1528-1534(1991)



J A W W A 84 98-104 (1992)





pp 48-58






(1994)






i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5




E COLI

1247


207-214 (1991)


2684 (1988)

Kampfer P

Rauhoff









foods




1476-1481 (1987)


59 786-790(1995)



M W at Sci T ec h 2 5 93-103 (1992)




Rice


(1990)







1248 ALONSO ET AL


(1993)


2236-2243 (1996)


(1976)



e c e i v e d

December

22 1997


i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5











E coli





(APHA 1995)






+

GUS ) and the




+

GUS)






i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5



1232 ALONSO ET AL


Sampling












K coli




Bacterial Strains














TABLE 1



1233

Test Strain






Klebsiella oxytoca

Klebsiella ozaenae




Escherichia coli

Hafnia alvei


Serratia marcescens

Serratia rubiadea

Cedecea davisae

Kluyvera ascorbata

Shigella flexneri

Shigella boydii

Shigella sonnei

Aeromonas hydrofila

Aeromonas caviae

Aeromonas media

A eromonas jandaei


Aeromonas trota


A

veronii bv veronii

Vibrio cholerae



Enterococcusfaecium

Salmonella derby


Salmonella london

No

684

194

858

857

140

860

851

856

863

401

678

157

867

159

868

842

861

585

583

413

398

838

4232

4228

4241

4255

4224

4257

557

108

184

410

IHMN

IHMN

IHMN

37degC

G

b

C

c

+ r

+ r

+ r

+ r

+ r

+ r

+ r

+ r

+ r

+ r

+ b

+ r

+ r

+ r

+ r

+ c

+ r

+ c

+ t

+ b

+ r

+ r

+ r

+ r

+ c

+ r

+ r

+ r

+ r

+ c

-

-

+ c

+ t

+ t

41

G ~

+

+

+

+

+

+

+

+

+

+

+

+

+

+

-

+

+

+

+

+

+

+

+

-

+

+

+

+

+

-

-

+

+

+

degC

r

r

r

r

r

r

r

r

c

r

b

r

r

t

r

-

r

c

t

b

r

r

r

r

-

r

r

r

r

c

-

-

c

t

t

445degC

G C

+ r

+ r

+ r

-

+ c

+ r

+ r

+ c

+ c

+ r

+ b

+ r

+ r

+ r

+ r

-

+ r

+ c

+ t

+ b

-

-

+ r

-

-

-

-

-

+ c

-

-

+ c

+ t

+ t


G


b

C


c=Colourless





1234 ALONSO ET AL












+



K coli

For














i t y ] a t 2 1 0 5 1 8 J u l y 2 0 1 5






(LAC

+


4








system (bioMerieux)















i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5



1236 ALONSO ET AL

TABLE 2

N o n - pound

coli

Fecal Coliforms

Klebsiella

spp

Enterobacter

spp and

Citrobacter

spp) KE C ) and

Escherich ia coli



Sampling

source

TR

EC-CCA

b

KEC-CCA

C

EKEC-CCA

d

F C - m F C

PI

EC-CCA

KEC-CCA

EKEC-CCA

FC-mFC

AP

E C - C C A

K E C - C C A

EKEC-CCA

FC-mFC

M l

EC-CCA

K E C - C C A

EKEC-CCA

FC-mFC

M2

EC-CCA

K E C - C C A

E K E C - C C A

FC-mFC

Mean

215

197

239

245

195

081

150

168

672

596

679

673

535

451

541

536

307

253

318

313

2h37deg-445degC

SD

151

163

156

167

062

025

080

071

020

014

019

019

124

112

122

125

093

083

091

098

Min

070

030

085

085

111

048

048

060

646

578

656

652

338

270

346

336

208

154

220

208

Max

408

420

445

461

258

108

259

262

700

623

705

703

672

570

676

671

448

370

454

462

Mean

287

264

237

241

179

060

161

161

671

582

676

673

532

439

537

536

300

240

311

307

445

SD

141

148

157

158

055

026

072

077

019

016

018

019

122

129

123

123

102

085

098

101

degC

Min

158

100

078

104

118

030

048

048

651

560

657

652

338

230

341

338

194

140

205

181

Max

426

420

453

448

250

095

250

256

699

611

703

704

669

604

678

668

457

378

463

458



and maximum (Max)



+


d




i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5







s
























1238

ALONSO ET AL

T A B L E 3



Sample

site

T R

P I

AP

M l

M 2

Parameters

EC37-KEC37

EC44-KEC44

EC37-KEC37

EC44-KEC44

EC37-KEC37

EC44-KEC44

EC37-KEC37

EC44-KEC44

EC37-KEC37

EC44-KEC44

R

099

098

069

064

068

040

099

099

099

098

P

lt001

lt001

N S

b

N S

N S

N S

lt001

lt001

lt001

lt001

Intercept

(a)

0340

0405

0571

1729

0977

3981

0419

1185

0257

0170

Slope

(b )

0916

0936

1701

0108

0963

0468

1093

0941

1112

0182

a












Aeromonas jandaei


i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5







+

G U S L A C

+

G U S

+

LAC



+

GUD species




+

GUS

4



15 L A C G U S

+


+


















i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5



24

ALONSO ET AL

TABLE 4

Num ber o f

coli

a nd N o n-E

coli



Reactions

Phenotype

LAC GUS

AP

C

M l

M 2

TR

PI

P2

Total

LAC

+

GUS

+

AP

M l

M 2

T R

PI

P2

Total

LACGUS

A P

M l

M 2

T R

P I

P2

Total

LAC GUS-

A P

M l

M2

TR

PI

P2

Total

Isolates

N o

37

31

42

41

31

11

193

24

29

48

48

52

11

212

2

9

0

3

2

0

16

17

23

15

36

46

29

166

E

N o

14

10

25

19

18

1

87

24

27

48

46

52

10

207

2

8

0

3

2

0

15

0

2

1

5

1

0

9

coli

( )

38

32

59

46

58

9

45

100

93

100

96

100

91

98

100

89

0

100

100

0

94

0

9

7

14

2

0

5

K E C

No

23

20

17

17

8

1

86

0

2

0

2

0

1

5

0

1

0

0

0

0

1

17

20

12

14

6

2

71

( )

62

68

41

42

26

9

45

0

7

0

4

0

9

2

0

11

0

0

0

0

6

100

87

80

39

13

7

43

Non

coliform

b

No

0

0

0

0

3

5

8

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

1

2

12

22

10

47

( )

0

0

0

0

10

46

4

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

4

13

33

48

34

28

N ot

identified

No

0

1

0

5

2

4

12

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

5

17

17

39

( )

0

3

0

12

6

36

6

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

14

37

59

24

KEC


b


Sampling sites



i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5



TAB LE 5


O

m

sect

o

L A C

+

G U S

N o

L A C

+


4

N o L A C G U S -

No


Klebsiella oxytoca

K Pneuntoniae

Citrobacterfreundii

C Amalonaticus

Escherichia coli

4

2

2

6

1

6

pound co

Cfreundii

8

1

ot l

E co

21

Pseudomonas

spp

P fluorescens

Acinetobacter

spp

Flavobacterium

spp

Proteus

spp

Salmonella typhi

Citrobacterfreundii

C amalonaticus

Klebsiella oxytoca

K pneumoniae


E agglomerans

E sakazakii

Escherichia coli

4

1

1

1

1

1

8

1

4

2

3

1

1

4

33

a

LA C

+


bull LAC GU S

+


l A C

G US

+


d





1242 ALONSO ET AL



1991










sample

itself


+

G U S

+










red colonies (LAC

+



7



i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5




G U S

+

colony and 5 LAC

+

GUS

+
























i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5



1244 ALONSO ET AL

When LAC

+

GUS LAC

+

GUS

+







LAC

+





+








1992)





non-salt adapted

E coli









TABLEOacute


K coli


Sampling

sites

AP

M l

M2

TR

PI

P2

N o of

strains

24

30

53

46

55

11

Thermo-

tolerant

No

24

28

51

42

51

9

100

93

96

91

93

82

Thermo-

sensitive

b

No

0

2

2

4

3

12

0

7

4

9

5

5

Indol-

No

0

2

3

2

1

9

0

7

6

4

2

4


k






REFERENCES


315(1996)



1-117


Bej


(1991)


i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5



1246 ALONSO ET AL


M icrobiol 59 3534-3544 (1993)




336-344 (1992)




Microbiol 57 1528-1534(1991)



J A W W A 84 98-104 (1992)





pp 48-58






(1994)






i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5




E COLI

1247


207-214 (1991)


2684 (1988)

Kampfer P

Rauhoff









foods




1476-1481 (1987)


59 786-790(1995)



M W at Sci T ec h 2 5 93-103 (1992)




Rice


(1990)







1248 ALONSO ET AL


(1993)


2236-2243 (1996)


(1976)



e c e i v e d

December

22 1997


i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5



1232 ALONSO ET AL


Sampling












K coli




Bacterial Strains














TABLE 1



1233

Test Strain






Klebsiella oxytoca

Klebsiella ozaenae




Escherichia coli

Hafnia alvei


Serratia marcescens

Serratia rubiadea

Cedecea davisae

Kluyvera ascorbata

Shigella flexneri

Shigella boydii

Shigella sonnei

Aeromonas hydrofila

Aeromonas caviae

Aeromonas media

A eromonas jandaei


Aeromonas trota


A

veronii bv veronii

Vibrio cholerae



Enterococcusfaecium

Salmonella derby


Salmonella london

No

684

194

858

857

140

860

851

856

863

401

678

157

867

159

868

842

861

585

583

413

398

838

4232

4228

4241

4255

4224

4257

557

108

184

410

IHMN

IHMN

IHMN

37degC

G

b

C

c

+ r

+ r

+ r

+ r

+ r

+ r

+ r

+ r

+ r

+ r

+ b

+ r

+ r

+ r

+ r

+ c

+ r

+ c

+ t

+ b

+ r

+ r

+ r

+ r

+ c

+ r

+ r

+ r

+ r

+ c

-

-

+ c

+ t

+ t

41

G ~

+

+

+

+

+

+

+

+

+

+

+

+

+

+

-

+

+

+

+

+

+

+

+

-

+

+

+

+

+

-

-

+

+

+

degC

r

r

r

r

r

r

r

r

c

r

b

r

r

t

r

-

r

c

t

b

r

r

r

r

-

r

r

r

r

c

-

-

c

t

t

445degC

G C

+ r

+ r

+ r

-

+ c

+ r

+ r

+ c

+ c

+ r

+ b

+ r

+ r

+ r

+ r

-

+ r

+ c

+ t

+ b

-

-

+ r

-

-

-

-

-

+ c

-

-

+ c

+ t

+ t


G


b

C


c=Colourless





1234 ALONSO ET AL












+



K coli

For














i t y ] a t 2 1 0 5 1 8 J u l y 2 0 1 5






(LAC

+


4








system (bioMerieux)















i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5



1236 ALONSO ET AL

TABLE 2

N o n - pound

coli

Fecal Coliforms

Klebsiella

spp

Enterobacter

spp and

Citrobacter

spp) KE C ) and

Escherich ia coli



Sampling

source

TR

EC-CCA

b

KEC-CCA

C

EKEC-CCA

d

F C - m F C

PI

EC-CCA

KEC-CCA

EKEC-CCA

FC-mFC

AP

E C - C C A

K E C - C C A

EKEC-CCA

FC-mFC

M l

EC-CCA

K E C - C C A

EKEC-CCA

FC-mFC

M2

EC-CCA

K E C - C C A

E K E C - C C A

FC-mFC

Mean

215

197

239

245

195

081

150

168

672

596

679

673

535

451

541

536

307

253

318

313

2h37deg-445degC

SD

151

163

156

167

062

025

080

071

020

014

019

019

124

112

122

125

093

083

091

098

Min

070

030

085

085

111

048

048

060

646

578

656

652

338

270

346

336

208

154

220

208

Max

408

420

445

461

258

108

259

262

700

623

705

703

672

570

676

671

448

370

454

462

Mean

287

264

237

241

179

060

161

161

671

582

676

673

532

439

537

536

300

240

311

307

445

SD

141

148

157

158

055

026

072

077

019

016

018

019

122

129

123

123

102

085

098

101

degC

Min

158

100

078

104

118

030

048

048

651

560

657

652

338

230

341

338

194

140

205

181

Max

426

420

453

448

250

095

250

256

699

611

703

704

669

604

678

668

457

378

463

458



and maximum (Max)



+


d




i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5







s
























1238

ALONSO ET AL

T A B L E 3



Sample

site

T R

P I

AP

M l

M 2

Parameters

EC37-KEC37

EC44-KEC44

EC37-KEC37

EC44-KEC44

EC37-KEC37

EC44-KEC44

EC37-KEC37

EC44-KEC44

EC37-KEC37

EC44-KEC44

R

099

098

069

064

068

040

099

099

099

098

P

lt001

lt001

N S

b

N S

N S

N S

lt001

lt001

lt001

lt001

Intercept

(a)

0340

0405

0571

1729

0977

3981

0419

1185

0257

0170

Slope

(b )

0916

0936

1701

0108

0963

0468

1093

0941

1112

0182

a












Aeromonas jandaei


i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5







+

G U S L A C

+

G U S

+

LAC



+

GUD species




+

GUS

4



15 L A C G U S

+


+


















i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5



24

ALONSO ET AL

TABLE 4

Num ber o f

coli

a nd N o n-E

coli



Reactions

Phenotype

LAC GUS

AP

C

M l

M 2

TR

PI

P2

Total

LAC

+

GUS

+

AP

M l

M 2

T R

PI

P2

Total

LACGUS

A P

M l

M 2

T R

P I

P2

Total

LAC GUS-

A P

M l

M2

TR

PI

P2

Total

Isolates

N o

37

31

42

41

31

11

193

24

29

48

48

52

11

212

2

9

0

3

2

0

16

17

23

15

36

46

29

166

E

N o

14

10

25

19

18

1

87

24

27

48

46

52

10

207

2

8

0

3

2

0

15

0

2

1

5

1

0

9

coli

( )

38

32

59

46

58

9

45

100

93

100

96

100

91

98

100

89

0

100

100

0

94

0

9

7

14

2

0

5

K E C

No

23

20

17

17

8

1

86

0

2

0

2

0

1

5

0

1

0

0

0

0

1

17

20

12

14

6

2

71

( )

62

68

41

42

26

9

45

0

7

0

4

0

9

2

0

11

0

0

0

0

6

100

87

80

39

13

7

43

Non

coliform

b

No

0

0

0

0

3

5

8

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

1

2

12

22

10

47

( )

0

0

0

0

10

46

4

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

4

13

33

48

34

28

N ot

identified

No

0

1

0

5

2

4

12

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

5

17

17

39

( )

0

3

0

12

6

36

6

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

14

37

59

24

KEC


b


Sampling sites



i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5



TAB LE 5


O

m

sect

o

L A C

+

G U S

N o

L A C

+


4

N o L A C G U S -

No


Klebsiella oxytoca

K Pneuntoniae

Citrobacterfreundii

C Amalonaticus

Escherichia coli

4

2

2

6

1

6

pound co

Cfreundii

8

1

ot l

E co

21

Pseudomonas

spp

P fluorescens

Acinetobacter

spp

Flavobacterium

spp

Proteus

spp

Salmonella typhi

Citrobacterfreundii

C amalonaticus

Klebsiella oxytoca

K pneumoniae


E agglomerans

E sakazakii

Escherichia coli

4

1

1

1

1

1

8

1

4

2

3

1

1

4

33

a

LA C

+


bull LAC GU S

+


l A C

G US

+


d





1242 ALONSO ET AL



1991










sample

itself


+

G U S

+










red colonies (LAC

+



7



i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5




G U S

+

colony and 5 LAC

+

GUS

+
























i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5



1244 ALONSO ET AL

When LAC

+

GUS LAC

+

GUS

+







LAC

+





+








1992)





non-salt adapted

E coli









TABLEOacute


K coli


Sampling

sites

AP

M l

M2

TR

PI

P2

N o of

strains

24

30

53

46

55

11

Thermo-

tolerant

No

24

28

51

42

51

9

100

93

96

91

93

82

Thermo-

sensitive

b

No

0

2

2

4

3

12

0

7

4

9

5

5

Indol-

No

0

2

3

2

1

9

0

7

6

4

2

4


k






REFERENCES


315(1996)



1-117


Bej


(1991)


i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5



1246 ALONSO ET AL


M icrobiol 59 3534-3544 (1993)




336-344 (1992)




Microbiol 57 1528-1534(1991)



J A W W A 84 98-104 (1992)





pp 48-58






(1994)






i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5




E COLI

1247


207-214 (1991)


2684 (1988)

Kampfer P

Rauhoff









foods




1476-1481 (1987)


59 786-790(1995)



M W at Sci T ec h 2 5 93-103 (1992)




Rice


(1990)







1248 ALONSO ET AL


(1993)


2236-2243 (1996)


(1976)



e c e i v e d

December

22 1997


i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5




TABLE 1



1233

Test Strain






Klebsiella oxytoca

Klebsiella ozaenae




Escherichia coli

Hafnia alvei


Serratia marcescens

Serratia rubiadea

Cedecea davisae

Kluyvera ascorbata

Shigella flexneri

Shigella boydii

Shigella sonnei

Aeromonas hydrofila

Aeromonas caviae

Aeromonas media

A eromonas jandaei


Aeromonas trota


A

veronii bv veronii

Vibrio cholerae



Enterococcusfaecium

Salmonella derby


Salmonella london

No

684

194

858

857

140

860

851

856

863

401

678

157

867

159

868

842

861

585

583

413

398

838

4232

4228

4241

4255

4224

4257

557

108

184

410

IHMN

IHMN

IHMN

37degC

G

b

C

c

+ r

+ r

+ r

+ r

+ r

+ r

+ r

+ r

+ r

+ r

+ b

+ r

+ r

+ r

+ r

+ c

+ r

+ c

+ t

+ b

+ r

+ r

+ r

+ r

+ c

+ r

+ r

+ r

+ r

+ c

-

-

+ c

+ t

+ t

41

G ~

+

+

+

+

+

+

+

+

+

+

+

+

+

+

-

+

+

+

+

+

+

+

+

-

+

+

+

+

+

-

-

+

+

+

degC

r

r

r

r

r

r

r

r

c

r

b

r

r

t

r

-

r

c

t

b

r

r

r

r

-

r

r

r

r

c

-

-

c

t

t

445degC

G C

+ r

+ r

+ r

-

+ c

+ r

+ r

+ c

+ c

+ r

+ b

+ r

+ r

+ r

+ r

-

+ r

+ c

+ t

+ b

-

-

+ r

-

-

-

-

-

+ c

-

-

+ c

+ t

+ t


G


b

C


c=Colourless





1234 ALONSO ET AL












+



K coli

For














i t y ] a t 2 1 0 5 1 8 J u l y 2 0 1 5






(LAC

+


4








system (bioMerieux)















i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5



1236 ALONSO ET AL

TABLE 2

N o n - pound

coli

Fecal Coliforms

Klebsiella

spp

Enterobacter

spp and

Citrobacter

spp) KE C ) and

Escherich ia coli



Sampling

source

TR

EC-CCA

b

KEC-CCA

C

EKEC-CCA

d

F C - m F C

PI

EC-CCA

KEC-CCA

EKEC-CCA

FC-mFC

AP

E C - C C A

K E C - C C A

EKEC-CCA

FC-mFC

M l

EC-CCA

K E C - C C A

EKEC-CCA

FC-mFC

M2

EC-CCA

K E C - C C A

E K E C - C C A

FC-mFC

Mean

215

197

239

245

195

081

150

168

672

596

679

673

535

451

541

536

307

253

318

313

2h37deg-445degC

SD

151

163

156

167

062

025

080

071

020

014

019

019

124

112

122

125

093

083

091

098

Min

070

030

085

085

111

048

048

060

646

578

656

652

338

270

346

336

208

154

220

208

Max

408

420

445

461

258

108

259

262

700

623

705

703

672

570

676

671

448

370

454

462

Mean

287

264

237

241

179

060

161

161

671

582

676

673

532

439

537

536

300

240

311

307

445

SD

141

148

157

158

055

026

072

077

019

016

018

019

122

129

123

123

102

085

098

101

degC

Min

158

100

078

104

118

030

048

048

651

560

657

652

338

230

341

338

194

140

205

181

Max

426

420

453

448

250

095

250

256

699

611

703

704

669

604

678

668

457

378

463

458



and maximum (Max)



+


d




i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5







s
























1238

ALONSO ET AL

T A B L E 3



Sample

site

T R

P I

AP

M l

M 2

Parameters

EC37-KEC37

EC44-KEC44

EC37-KEC37

EC44-KEC44

EC37-KEC37

EC44-KEC44

EC37-KEC37

EC44-KEC44

EC37-KEC37

EC44-KEC44

R

099

098

069

064

068

040

099

099

099

098

P

lt001

lt001

N S

b

N S

N S

N S

lt001

lt001

lt001

lt001

Intercept

(a)

0340

0405

0571

1729

0977

3981

0419

1185

0257

0170

Slope

(b )

0916

0936

1701

0108

0963

0468

1093

0941

1112

0182

a












Aeromonas jandaei


i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5







+

G U S L A C

+

G U S

+

LAC



+

GUD species




+

GUS

4



15 L A C G U S

+


+


















i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5



24

ALONSO ET AL

TABLE 4

Num ber o f

coli

a nd N o n-E

coli



Reactions

Phenotype

LAC GUS

AP

C

M l

M 2

TR

PI

P2

Total

LAC

+

GUS

+

AP

M l

M 2

T R

PI

P2

Total

LACGUS

A P

M l

M 2

T R

P I

P2

Total

LAC GUS-

A P

M l

M2

TR

PI

P2

Total

Isolates

N o

37

31

42

41

31

11

193

24

29

48

48

52

11

212

2

9

0

3

2

0

16

17

23

15

36

46

29

166

E

N o

14

10

25

19

18

1

87

24

27

48

46

52

10

207

2

8

0

3

2

0

15

0

2

1

5

1

0

9

coli

( )

38

32

59

46

58

9

45

100

93

100

96

100

91

98

100

89

0

100

100

0

94

0

9

7

14

2

0

5

K E C

No

23

20

17

17

8

1

86

0

2

0

2

0

1

5

0

1

0

0

0

0

1

17

20

12

14

6

2

71

( )

62

68

41

42

26

9

45

0

7

0

4

0

9

2

0

11

0

0

0

0

6

100

87

80

39

13

7

43

Non

coliform

b

No

0

0

0

0

3

5

8

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

1

2

12

22

10

47

( )

0

0

0

0

10

46

4

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

4

13

33

48

34

28

N ot

identified

No

0

1

0

5

2

4

12

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

5

17

17

39

( )

0

3

0

12

6

36

6

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

14

37

59

24

KEC


b


Sampling sites



i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5



TAB LE 5


O

m

sect

o

L A C

+

G U S

N o

L A C

+


4

N o L A C G U S -

No


Klebsiella oxytoca

K Pneuntoniae

Citrobacterfreundii

C Amalonaticus

Escherichia coli

4

2

2

6

1

6

pound co

Cfreundii

8

1

ot l

E co

21

Pseudomonas

spp

P fluorescens

Acinetobacter

spp

Flavobacterium

spp

Proteus

spp

Salmonella typhi

Citrobacterfreundii

C amalonaticus

Klebsiella oxytoca

K pneumoniae


E agglomerans

E sakazakii

Escherichia coli

4

1

1

1

1

1

8

1

4

2

3

1

1

4

33

a

LA C

+


bull LAC GU S

+


l A C

G US

+


d





1242 ALONSO ET AL



1991










sample

itself


+

G U S

+










red colonies (LAC

+



7



i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5




G U S

+

colony and 5 LAC

+

GUS

+
























i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5



1244 ALONSO ET AL

When LAC

+

GUS LAC

+

GUS

+







LAC

+





+








1992)





non-salt adapted

E coli









TABLEOacute


K coli


Sampling

sites

AP

M l

M2

TR

PI

P2

N o of

strains

24

30

53

46

55

11

Thermo-

tolerant

No

24

28

51

42

51

9

100

93

96

91

93

82

Thermo-

sensitive

b

No

0

2

2

4

3

12

0

7

4

9

5

5

Indol-

No

0

2

3

2

1

9

0

7

6

4

2

4


k






REFERENCES


315(1996)



1-117


Bej


(1991)


i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5



1246 ALONSO ET AL


M icrobiol 59 3534-3544 (1993)




336-344 (1992)




Microbiol 57 1528-1534(1991)



J A W W A 84 98-104 (1992)





pp 48-58






(1994)






i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5




E COLI

1247


207-214 (1991)


2684 (1988)

Kampfer P

Rauhoff









foods




1476-1481 (1987)


59 786-790(1995)



M W at Sci T ec h 2 5 93-103 (1992)




Rice


(1990)







1248 ALONSO ET AL


(1993)


2236-2243 (1996)


(1976)



e c e i v e d

December

22 1997


i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5



1234 ALONSO ET AL












+



K coli

For














i t y ] a t 2 1 0 5 1 8 J u l y 2 0 1 5






(LAC

+


4








system (bioMerieux)















i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5



1236 ALONSO ET AL

TABLE 2

N o n - pound

coli

Fecal Coliforms

Klebsiella

spp

Enterobacter

spp and

Citrobacter

spp) KE C ) and

Escherich ia coli



Sampling

source

TR

EC-CCA

b

KEC-CCA

C

EKEC-CCA

d

F C - m F C

PI

EC-CCA

KEC-CCA

EKEC-CCA

FC-mFC

AP

E C - C C A

K E C - C C A

EKEC-CCA

FC-mFC

M l

EC-CCA

K E C - C C A

EKEC-CCA

FC-mFC

M2

EC-CCA

K E C - C C A

E K E C - C C A

FC-mFC

Mean

215

197

239

245

195

081

150

168

672

596

679

673

535

451

541

536

307

253

318

313

2h37deg-445degC

SD

151

163

156

167

062

025

080

071

020

014

019

019

124

112

122

125

093

083

091

098

Min

070

030

085

085

111

048

048

060

646

578

656

652

338

270

346

336

208

154

220

208

Max

408

420

445

461

258

108

259

262

700

623

705

703

672

570

676

671

448

370

454

462

Mean

287

264

237

241

179

060

161

161

671

582

676

673

532

439

537

536

300

240

311

307

445

SD

141

148

157

158

055

026

072

077

019

016

018

019

122

129

123

123

102

085

098

101

degC

Min

158

100

078

104

118

030

048

048

651

560

657

652

338

230

341

338

194

140

205

181

Max

426

420

453

448

250

095

250

256

699

611

703

704

669

604

678

668

457

378

463

458



and maximum (Max)



+


d




i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5







s
























1238

ALONSO ET AL

T A B L E 3



Sample

site

T R

P I

AP

M l

M 2

Parameters

EC37-KEC37

EC44-KEC44

EC37-KEC37

EC44-KEC44

EC37-KEC37

EC44-KEC44

EC37-KEC37

EC44-KEC44

EC37-KEC37

EC44-KEC44

R

099

098

069

064

068

040

099

099

099

098

P

lt001

lt001

N S

b

N S

N S

N S

lt001

lt001

lt001

lt001

Intercept

(a)

0340

0405

0571

1729

0977

3981

0419

1185

0257

0170

Slope

(b )

0916

0936

1701

0108

0963

0468

1093

0941

1112

0182

a












Aeromonas jandaei


i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5







+

G U S L A C

+

G U S

+

LAC



+

GUD species




+

GUS

4



15 L A C G U S

+


+


















i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5



24

ALONSO ET AL

TABLE 4

Num ber o f

coli

a nd N o n-E

coli



Reactions

Phenotype

LAC GUS

AP

C

M l

M 2

TR

PI

P2

Total

LAC

+

GUS

+

AP

M l

M 2

T R

PI

P2

Total

LACGUS

A P

M l

M 2

T R

P I

P2

Total

LAC GUS-

A P

M l

M2

TR

PI

P2

Total

Isolates

N o

37

31

42

41

31

11

193

24

29

48

48

52

11

212

2

9

0

3

2

0

16

17

23

15

36

46

29

166

E

N o

14

10

25

19

18

1

87

24

27

48

46

52

10

207

2

8

0

3

2

0

15

0

2

1

5

1

0

9

coli

( )

38

32

59

46

58

9

45

100

93

100

96

100

91

98

100

89

0

100

100

0

94

0

9

7

14

2

0

5

K E C

No

23

20

17

17

8

1

86

0

2

0

2

0

1

5

0

1

0

0

0

0

1

17

20

12

14

6

2

71

( )

62

68

41

42

26

9

45

0

7

0

4

0

9

2

0

11

0

0

0

0

6

100

87

80

39

13

7

43

Non

coliform

b

No

0

0

0

0

3

5

8

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

1

2

12

22

10

47

( )

0

0

0

0

10

46

4

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

4

13

33

48

34

28

N ot

identified

No

0

1

0

5

2

4

12

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

5

17

17

39

( )

0

3

0

12

6

36

6

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

14

37

59

24

KEC


b


Sampling sites



i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5



TAB LE 5


O

m

sect

o

L A C

+

G U S

N o

L A C

+


4

N o L A C G U S -

No


Klebsiella oxytoca

K Pneuntoniae

Citrobacterfreundii

C Amalonaticus

Escherichia coli

4

2

2

6

1

6

pound co

Cfreundii

8

1

ot l

E co

21

Pseudomonas

spp

P fluorescens

Acinetobacter

spp

Flavobacterium

spp

Proteus

spp

Salmonella typhi

Citrobacterfreundii

C amalonaticus

Klebsiella oxytoca

K pneumoniae


E agglomerans

E sakazakii

Escherichia coli

4

1

1

1

1

1

8

1

4

2

3

1

1

4

33

a

LA C

+


bull LAC GU S

+


l A C

G US

+


d





1242 ALONSO ET AL



1991










sample

itself


+

G U S

+










red colonies (LAC

+



7



i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5




G U S

+

colony and 5 LAC

+

GUS

+
























i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5



1244 ALONSO ET AL

When LAC

+

GUS LAC

+

GUS

+







LAC

+





+








1992)





non-salt adapted

E coli









TABLEOacute


K coli


Sampling

sites

AP

M l

M2

TR

PI

P2

N o of

strains

24

30

53

46

55

11

Thermo-

tolerant

No

24

28

51

42

51

9

100

93

96

91

93

82

Thermo-

sensitive

b

No

0

2

2

4

3

12

0

7

4

9

5

5

Indol-

No

0

2

3

2

1

9

0

7

6

4

2

4


k






REFERENCES


315(1996)



1-117


Bej


(1991)


i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5



1246 ALONSO ET AL


M icrobiol 59 3534-3544 (1993)




336-344 (1992)




Microbiol 57 1528-1534(1991)



J A W W A 84 98-104 (1992)





pp 48-58






(1994)






i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5




E COLI

1247


207-214 (1991)


2684 (1988)

Kampfer P

Rauhoff









foods




1476-1481 (1987)


59 786-790(1995)



M W at Sci T ec h 2 5 93-103 (1992)




Rice


(1990)







1248 ALONSO ET AL


(1993)


2236-2243 (1996)


(1976)



e c e i v e d

December

22 1997


i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5






(LAC

+


4








system (bioMerieux)















i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5



1236 ALONSO ET AL

TABLE 2

N o n - pound

coli

Fecal Coliforms

Klebsiella

spp

Enterobacter

spp and

Citrobacter

spp) KE C ) and

Escherich ia coli



Sampling

source

TR

EC-CCA

b

KEC-CCA

C

EKEC-CCA

d

F C - m F C

PI

EC-CCA

KEC-CCA

EKEC-CCA

FC-mFC

AP

E C - C C A

K E C - C C A

EKEC-CCA

FC-mFC

M l

EC-CCA

K E C - C C A

EKEC-CCA

FC-mFC

M2

EC-CCA

K E C - C C A

E K E C - C C A

FC-mFC

Mean

215

197

239

245

195

081

150

168

672

596

679

673

535

451

541

536

307

253

318

313

2h37deg-445degC

SD

151

163

156

167

062

025

080

071

020

014

019

019

124

112

122

125

093

083

091

098

Min

070

030

085

085

111

048

048

060

646

578

656

652

338

270

346

336

208

154

220

208

Max

408

420

445

461

258

108

259

262

700

623

705

703

672

570

676

671

448

370

454

462

Mean

287

264

237

241

179

060

161

161

671

582

676

673

532

439

537

536

300

240

311

307

445

SD

141

148

157

158

055

026

072

077

019

016

018

019

122

129

123

123

102

085

098

101

degC

Min

158

100

078

104

118

030

048

048

651

560

657

652

338

230

341

338

194

140

205

181

Max

426

420

453

448

250

095

250

256

699

611

703

704

669

604

678

668

457

378

463

458



and maximum (Max)



+


d




i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5







s
























1238

ALONSO ET AL

T A B L E 3



Sample

site

T R

P I

AP

M l

M 2

Parameters

EC37-KEC37

EC44-KEC44

EC37-KEC37

EC44-KEC44

EC37-KEC37

EC44-KEC44

EC37-KEC37

EC44-KEC44

EC37-KEC37

EC44-KEC44

R

099

098

069

064

068

040

099

099

099

098

P

lt001

lt001

N S

b

N S

N S

N S

lt001

lt001

lt001

lt001

Intercept

(a)

0340

0405

0571

1729

0977

3981

0419

1185

0257

0170

Slope

(b )

0916

0936

1701

0108

0963

0468

1093

0941

1112

0182

a












Aeromonas jandaei


i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5







+

G U S L A C

+

G U S

+

LAC



+

GUD species




+

GUS

4



15 L A C G U S

+


+


















i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5



24

ALONSO ET AL

TABLE 4

Num ber o f

coli

a nd N o n-E

coli



Reactions

Phenotype

LAC GUS

AP

C

M l

M 2

TR

PI

P2

Total

LAC

+

GUS

+

AP

M l

M 2

T R

PI

P2

Total

LACGUS

A P

M l

M 2

T R

P I

P2

Total

LAC GUS-

A P

M l

M2

TR

PI

P2

Total

Isolates

N o

37

31

42

41

31

11

193

24

29

48

48

52

11

212

2

9

0

3

2

0

16

17

23

15

36

46

29

166

E

N o

14

10

25

19

18

1

87

24

27

48

46

52

10

207

2

8

0

3

2

0

15

0

2

1

5

1

0

9

coli

( )

38

32

59

46

58

9

45

100

93

100

96

100

91

98

100

89

0

100

100

0

94

0

9

7

14

2

0

5

K E C

No

23

20

17

17

8

1

86

0

2

0

2

0

1

5

0

1

0

0

0

0

1

17

20

12

14

6

2

71

( )

62

68

41

42

26

9

45

0

7

0

4

0

9

2

0

11

0

0

0

0

6

100

87

80

39

13

7

43

Non

coliform

b

No

0

0

0

0

3

5

8

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

1

2

12

22

10

47

( )

0

0

0

0

10

46

4

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

4

13

33

48

34

28

N ot

identified

No

0

1

0

5

2

4

12

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

5

17

17

39

( )

0

3

0

12

6

36

6

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

14

37

59

24

KEC


b


Sampling sites



i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5



TAB LE 5


O

m

sect

o

L A C

+

G U S

N o

L A C

+


4

N o L A C G U S -

No


Klebsiella oxytoca

K Pneuntoniae

Citrobacterfreundii

C Amalonaticus

Escherichia coli

4

2

2

6

1

6

pound co

Cfreundii

8

1

ot l

E co

21

Pseudomonas

spp

P fluorescens

Acinetobacter

spp

Flavobacterium

spp

Proteus

spp

Salmonella typhi

Citrobacterfreundii

C amalonaticus

Klebsiella oxytoca

K pneumoniae


E agglomerans

E sakazakii

Escherichia coli

4

1

1

1

1

1

8

1

4

2

3

1

1

4

33

a

LA C

+


bull LAC GU S

+


l A C

G US

+


d





1242 ALONSO ET AL



1991










sample

itself


+

G U S

+










red colonies (LAC

+



7



i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5




G U S

+

colony and 5 LAC

+

GUS

+
























i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5



1244 ALONSO ET AL

When LAC

+

GUS LAC

+

GUS

+







LAC

+





+








1992)





non-salt adapted

E coli









TABLEOacute


K coli


Sampling

sites

AP

M l

M2

TR

PI

P2

N o of

strains

24

30

53

46

55

11

Thermo-

tolerant

No

24

28

51

42

51

9

100

93

96

91

93

82

Thermo-

sensitive

b

No

0

2

2

4

3

12

0

7

4

9

5

5

Indol-

No

0

2

3

2

1

9

0

7

6

4

2

4


k






REFERENCES


315(1996)



1-117


Bej


(1991)


i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5



1246 ALONSO ET AL


M icrobiol 59 3534-3544 (1993)




336-344 (1992)




Microbiol 57 1528-1534(1991)



J A W W A 84 98-104 (1992)





pp 48-58






(1994)






i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5




E COLI

1247


207-214 (1991)


2684 (1988)

Kampfer P

Rauhoff









foods




1476-1481 (1987)


59 786-790(1995)



M W at Sci T ec h 2 5 93-103 (1992)




Rice


(1990)







1248 ALONSO ET AL


(1993)


2236-2243 (1996)


(1976)



e c e i v e d

December

22 1997


i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5



1236 ALONSO ET AL

TABLE 2

N o n - pound

coli

Fecal Coliforms

Klebsiella

spp

Enterobacter

spp and

Citrobacter

spp) KE C ) and

Escherich ia coli



Sampling

source

TR

EC-CCA

b

KEC-CCA

C

EKEC-CCA

d

F C - m F C

PI

EC-CCA

KEC-CCA

EKEC-CCA

FC-mFC

AP

E C - C C A

K E C - C C A

EKEC-CCA

FC-mFC

M l

EC-CCA

K E C - C C A

EKEC-CCA

FC-mFC

M2

EC-CCA

K E C - C C A

E K E C - C C A

FC-mFC

Mean

215

197

239

245

195

081

150

168

672

596

679

673

535

451

541

536

307

253

318

313

2h37deg-445degC

SD

151

163

156

167

062

025

080

071

020

014

019

019

124

112

122

125

093

083

091

098

Min

070

030

085

085

111

048

048

060

646

578

656

652

338

270

346

336

208

154

220

208

Max

408

420

445

461

258

108

259

262

700

623

705

703

672

570

676

671

448

370

454

462

Mean

287

264

237

241

179

060

161

161

671

582

676

673

532

439

537

536

300

240

311

307

445

SD

141

148

157

158

055

026

072

077

019

016

018

019

122

129

123

123

102

085

098

101

degC

Min

158

100

078

104

118

030

048

048

651

560

657

652

338

230

341

338

194

140

205

181

Max

426

420

453

448

250

095

250

256

699

611

703

704

669

604

678

668

457

378

463

458



and maximum (Max)



+


d




i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5







s
























1238

ALONSO ET AL

T A B L E 3



Sample

site

T R

P I

AP

M l

M 2

Parameters

EC37-KEC37

EC44-KEC44

EC37-KEC37

EC44-KEC44

EC37-KEC37

EC44-KEC44

EC37-KEC37

EC44-KEC44

EC37-KEC37

EC44-KEC44

R

099

098

069

064

068

040

099

099

099

098

P

lt001

lt001

N S

b

N S

N S

N S

lt001

lt001

lt001

lt001

Intercept

(a)

0340

0405

0571

1729

0977

3981

0419

1185

0257

0170

Slope

(b )

0916

0936

1701

0108

0963

0468

1093

0941

1112

0182

a












Aeromonas jandaei


i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5







+

G U S L A C

+

G U S

+

LAC



+

GUD species




+

GUS

4



15 L A C G U S

+


+


















i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5



24

ALONSO ET AL

TABLE 4

Num ber o f

coli

a nd N o n-E

coli



Reactions

Phenotype

LAC GUS

AP

C

M l

M 2

TR

PI

P2

Total

LAC

+

GUS

+

AP

M l

M 2

T R

PI

P2

Total

LACGUS

A P

M l

M 2

T R

P I

P2

Total

LAC GUS-

A P

M l

M2

TR

PI

P2

Total

Isolates

N o

37

31

42

41

31

11

193

24

29

48

48

52

11

212

2

9

0

3

2

0

16

17

23

15

36

46

29

166

E

N o

14

10

25

19

18

1

87

24

27

48

46

52

10

207

2

8

0

3

2

0

15

0

2

1

5

1

0

9

coli

( )

38

32

59

46

58

9

45

100

93

100

96

100

91

98

100

89

0

100

100

0

94

0

9

7

14

2

0

5

K E C

No

23

20

17

17

8

1

86

0

2

0

2

0

1

5

0

1

0

0

0

0

1

17

20

12

14

6

2

71

( )

62

68

41

42

26

9

45

0

7

0

4

0

9

2

0

11

0

0

0

0

6

100

87

80

39

13

7

43

Non

coliform

b

No

0

0

0

0

3

5

8

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

1

2

12

22

10

47

( )

0

0

0

0

10

46

4

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

4

13

33

48

34

28

N ot

identified

No

0

1

0

5

2

4

12

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

5

17

17

39

( )

0

3

0

12

6

36

6

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

14

37

59

24

KEC


b


Sampling sites



i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5



TAB LE 5


O

m

sect

o

L A C

+

G U S

N o

L A C

+


4

N o L A C G U S -

No


Klebsiella oxytoca

K Pneuntoniae

Citrobacterfreundii

C Amalonaticus

Escherichia coli

4

2

2

6

1

6

pound co

Cfreundii

8

1

ot l

E co

21

Pseudomonas

spp

P fluorescens

Acinetobacter

spp

Flavobacterium

spp

Proteus

spp

Salmonella typhi

Citrobacterfreundii

C amalonaticus

Klebsiella oxytoca

K pneumoniae


E agglomerans

E sakazakii

Escherichia coli

4

1

1

1

1

1

8

1

4

2

3

1

1

4

33

a

LA C

+


bull LAC GU S

+


l A C

G US

+


d





1242 ALONSO ET AL



1991










sample

itself


+

G U S

+










red colonies (LAC

+



7



i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5




G U S

+

colony and 5 LAC

+

GUS

+
























i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5



1244 ALONSO ET AL

When LAC

+

GUS LAC

+

GUS

+







LAC

+





+








1992)





non-salt adapted

E coli









TABLEOacute


K coli


Sampling

sites

AP

M l

M2

TR

PI

P2

N o of

strains

24

30

53

46

55

11

Thermo-

tolerant

No

24

28

51

42

51

9

100

93

96

91

93

82

Thermo-

sensitive

b

No

0

2

2

4

3

12

0

7

4

9

5

5

Indol-

No

0

2

3

2

1

9

0

7

6

4

2

4


k






REFERENCES


315(1996)



1-117


Bej


(1991)


i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5



1246 ALONSO ET AL


M icrobiol 59 3534-3544 (1993)




336-344 (1992)




Microbiol 57 1528-1534(1991)



J A W W A 84 98-104 (1992)





pp 48-58






(1994)






i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5




E COLI

1247


207-214 (1991)


2684 (1988)

Kampfer P

Rauhoff









foods




1476-1481 (1987)


59 786-790(1995)



M W at Sci T ec h 2 5 93-103 (1992)




Rice


(1990)







1248 ALONSO ET AL


(1993)


2236-2243 (1996)


(1976)



e c e i v e d

December

22 1997


i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5







s
























1238

ALONSO ET AL

T A B L E 3



Sample

site

T R

P I

AP

M l

M 2

Parameters

EC37-KEC37

EC44-KEC44

EC37-KEC37

EC44-KEC44

EC37-KEC37

EC44-KEC44

EC37-KEC37

EC44-KEC44

EC37-KEC37

EC44-KEC44

R

099

098

069

064

068

040

099

099

099

098

P

lt001

lt001

N S

b

N S

N S

N S

lt001

lt001

lt001

lt001

Intercept

(a)

0340

0405

0571

1729

0977

3981

0419

1185

0257

0170

Slope

(b )

0916

0936

1701

0108

0963

0468

1093

0941

1112

0182

a












Aeromonas jandaei


i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5







+

G U S L A C

+

G U S

+

LAC



+

GUD species




+

GUS

4



15 L A C G U S

+


+


















i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5



24

ALONSO ET AL

TABLE 4

Num ber o f

coli

a nd N o n-E

coli



Reactions

Phenotype

LAC GUS

AP

C

M l

M 2

TR

PI

P2

Total

LAC

+

GUS

+

AP

M l

M 2

T R

PI

P2

Total

LACGUS

A P

M l

M 2

T R

P I

P2

Total

LAC GUS-

A P

M l

M2

TR

PI

P2

Total

Isolates

N o

37

31

42

41

31

11

193

24

29

48

48

52

11

212

2

9

0

3

2

0

16

17

23

15

36

46

29

166

E

N o

14

10

25

19

18

1

87

24

27

48

46

52

10

207

2

8

0

3

2

0

15

0

2

1

5

1

0

9

coli

( )

38

32

59

46

58

9

45

100

93

100

96

100

91

98

100

89

0

100

100

0

94

0

9

7

14

2

0

5

K E C

No

23

20

17

17

8

1

86

0

2

0

2

0

1

5

0

1

0

0

0

0

1

17

20

12

14

6

2

71

( )

62

68

41

42

26

9

45

0

7

0

4

0

9

2

0

11

0

0

0

0

6

100

87

80

39

13

7

43

Non

coliform

b

No

0

0

0

0

3

5

8

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

1

2

12

22

10

47

( )

0

0

0

0

10

46

4

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

4

13

33

48

34

28

N ot

identified

No

0

1

0

5

2

4

12

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

5

17

17

39

( )

0

3

0

12

6

36

6

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

14

37

59

24

KEC


b


Sampling sites



i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5



TAB LE 5


O

m

sect

o

L A C

+

G U S

N o

L A C

+


4

N o L A C G U S -

No


Klebsiella oxytoca

K Pneuntoniae

Citrobacterfreundii

C Amalonaticus

Escherichia coli

4

2

2

6

1

6

pound co

Cfreundii

8

1

ot l

E co

21

Pseudomonas

spp

P fluorescens

Acinetobacter

spp

Flavobacterium

spp

Proteus

spp

Salmonella typhi

Citrobacterfreundii

C amalonaticus

Klebsiella oxytoca

K pneumoniae


E agglomerans

E sakazakii

Escherichia coli

4

1

1

1

1

1

8

1

4

2

3

1

1

4

33

a

LA C

+


bull LAC GU S

+


l A C

G US

+


d





1242 ALONSO ET AL



1991










sample

itself


+

G U S

+










red colonies (LAC

+



7



i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5




G U S

+

colony and 5 LAC

+

GUS

+
























i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5



1244 ALONSO ET AL

When LAC

+

GUS LAC

+

GUS

+







LAC

+





+








1992)





non-salt adapted

E coli









TABLEOacute


K coli


Sampling

sites

AP

M l

M2

TR

PI

P2

N o of

strains

24

30

53

46

55

11

Thermo-

tolerant

No

24

28

51

42

51

9

100

93

96

91

93

82

Thermo-

sensitive

b

No

0

2

2

4

3

12

0

7

4

9

5

5

Indol-

No

0

2

3

2

1

9

0

7

6

4

2

4


k






REFERENCES


315(1996)



1-117


Bej


(1991)


i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5



1246 ALONSO ET AL


M icrobiol 59 3534-3544 (1993)




336-344 (1992)




Microbiol 57 1528-1534(1991)



J A W W A 84 98-104 (1992)





pp 48-58






(1994)






i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5




E COLI

1247


207-214 (1991)


2684 (1988)

Kampfer P

Rauhoff









foods




1476-1481 (1987)


59 786-790(1995)



M W at Sci T ec h 2 5 93-103 (1992)




Rice


(1990)







1248 ALONSO ET AL


(1993)


2236-2243 (1996)


(1976)



e c e i v e d

December

22 1997


i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5



1238

ALONSO ET AL

T A B L E 3



Sample

site

T R

P I

AP

M l

M 2

Parameters

EC37-KEC37

EC44-KEC44

EC37-KEC37

EC44-KEC44

EC37-KEC37

EC44-KEC44

EC37-KEC37

EC44-KEC44

EC37-KEC37

EC44-KEC44

R

099

098

069

064

068

040

099

099

099

098

P

lt001

lt001

N S

b

N S

N S

N S

lt001

lt001

lt001

lt001

Intercept

(a)

0340

0405

0571

1729

0977

3981

0419

1185

0257

0170

Slope

(b )

0916

0936

1701

0108

0963

0468

1093

0941

1112

0182

a












Aeromonas jandaei


i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5







+

G U S L A C

+

G U S

+

LAC



+

GUD species




+

GUS

4



15 L A C G U S

+


+


















i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5



24

ALONSO ET AL

TABLE 4

Num ber o f

coli

a nd N o n-E

coli



Reactions

Phenotype

LAC GUS

AP

C

M l

M 2

TR

PI

P2

Total

LAC

+

GUS

+

AP

M l

M 2

T R

PI

P2

Total

LACGUS

A P

M l

M 2

T R

P I

P2

Total

LAC GUS-

A P

M l

M2

TR

PI

P2

Total

Isolates

N o

37

31

42

41

31

11

193

24

29

48

48

52

11

212

2

9

0

3

2

0

16

17

23

15

36

46

29

166

E

N o

14

10

25

19

18

1

87

24

27

48

46

52

10

207

2

8

0

3

2

0

15

0

2

1

5

1

0

9

coli

( )

38

32

59

46

58

9

45

100

93

100

96

100

91

98

100

89

0

100

100

0

94

0

9

7

14

2

0

5

K E C

No

23

20

17

17

8

1

86

0

2

0

2

0

1

5

0

1

0

0

0

0

1

17

20

12

14

6

2

71

( )

62

68

41

42

26

9

45

0

7

0

4

0

9

2

0

11

0

0

0

0

6

100

87

80

39

13

7

43

Non

coliform

b

No

0

0

0

0

3

5

8

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

1

2

12

22

10

47

( )

0

0

0

0

10

46

4

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

4

13

33

48

34

28

N ot

identified

No

0

1

0

5

2

4

12

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

5

17

17

39

( )

0

3

0

12

6

36

6

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

14

37

59

24

KEC


b


Sampling sites



i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5



TAB LE 5


O

m

sect

o

L A C

+

G U S

N o

L A C

+


4

N o L A C G U S -

No


Klebsiella oxytoca

K Pneuntoniae

Citrobacterfreundii

C Amalonaticus

Escherichia coli

4

2

2

6

1

6

pound co

Cfreundii

8

1

ot l

E co

21

Pseudomonas

spp

P fluorescens

Acinetobacter

spp

Flavobacterium

spp

Proteus

spp

Salmonella typhi

Citrobacterfreundii

C amalonaticus

Klebsiella oxytoca

K pneumoniae


E agglomerans

E sakazakii

Escherichia coli

4

1

1

1

1

1

8

1

4

2

3

1

1

4

33

a

LA C

+


bull LAC GU S

+


l A C

G US

+


d





1242 ALONSO ET AL



1991










sample

itself


+

G U S

+










red colonies (LAC

+



7



i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5




G U S

+

colony and 5 LAC

+

GUS

+
























i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5



1244 ALONSO ET AL

When LAC

+

GUS LAC

+

GUS

+







LAC

+





+








1992)





non-salt adapted

E coli









TABLEOacute


K coli


Sampling

sites

AP

M l

M2

TR

PI

P2

N o of

strains

24

30

53

46

55

11

Thermo-

tolerant

No

24

28

51

42

51

9

100

93

96

91

93

82

Thermo-

sensitive

b

No

0

2

2

4

3

12

0

7

4

9

5

5

Indol-

No

0

2

3

2

1

9

0

7

6

4

2

4


k






REFERENCES


315(1996)



1-117


Bej


(1991)


i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5



1246 ALONSO ET AL


M icrobiol 59 3534-3544 (1993)




336-344 (1992)




Microbiol 57 1528-1534(1991)



J A W W A 84 98-104 (1992)





pp 48-58






(1994)






i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5




E COLI

1247


207-214 (1991)


2684 (1988)

Kampfer P

Rauhoff









foods




1476-1481 (1987)


59 786-790(1995)



M W at Sci T ec h 2 5 93-103 (1992)




Rice


(1990)







1248 ALONSO ET AL


(1993)


2236-2243 (1996)


(1976)



e c e i v e d

December

22 1997


i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5







+

G U S L A C

+

G U S

+

LAC



+

GUD species




+

GUS

4



15 L A C G U S

+


+


















i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5



24

ALONSO ET AL

TABLE 4

Num ber o f

coli

a nd N o n-E

coli



Reactions

Phenotype

LAC GUS

AP

C

M l

M 2

TR

PI

P2

Total

LAC

+

GUS

+

AP

M l

M 2

T R

PI

P2

Total

LACGUS

A P

M l

M 2

T R

P I

P2

Total

LAC GUS-

A P

M l

M2

TR

PI

P2

Total

Isolates

N o

37

31

42

41

31

11

193

24

29

48

48

52

11

212

2

9

0

3

2

0

16

17

23

15

36

46

29

166

E

N o

14

10

25

19

18

1

87

24

27

48

46

52

10

207

2

8

0

3

2

0

15

0

2

1

5

1

0

9

coli

( )

38

32

59

46

58

9

45

100

93

100

96

100

91

98

100

89

0

100

100

0

94

0

9

7

14

2

0

5

K E C

No

23

20

17

17

8

1

86

0

2

0

2

0

1

5

0

1

0

0

0

0

1

17

20

12

14

6

2

71

( )

62

68

41

42

26

9

45

0

7

0

4

0

9

2

0

11

0

0

0

0

6

100

87

80

39

13

7

43

Non

coliform

b

No

0

0

0

0

3

5

8

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

1

2

12

22

10

47

( )

0

0

0

0

10

46

4

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

4

13

33

48

34

28

N ot

identified

No

0

1

0

5

2

4

12

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

5

17

17

39

( )

0

3

0

12

6

36

6

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

14

37

59

24

KEC


b


Sampling sites



i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5



TAB LE 5


O

m

sect

o

L A C

+

G U S

N o

L A C

+


4

N o L A C G U S -

No


Klebsiella oxytoca

K Pneuntoniae

Citrobacterfreundii

C Amalonaticus

Escherichia coli

4

2

2

6

1

6

pound co

Cfreundii

8

1

ot l

E co

21

Pseudomonas

spp

P fluorescens

Acinetobacter

spp

Flavobacterium

spp

Proteus

spp

Salmonella typhi

Citrobacterfreundii

C amalonaticus

Klebsiella oxytoca

K pneumoniae


E agglomerans

E sakazakii

Escherichia coli

4

1

1

1

1

1

8

1

4

2

3

1

1

4

33

a

LA C

+


bull LAC GU S

+


l A C

G US

+


d





1242 ALONSO ET AL



1991










sample

itself


+

G U S

+










red colonies (LAC

+



7



i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5




G U S

+

colony and 5 LAC

+

GUS

+
























i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5



1244 ALONSO ET AL

When LAC

+

GUS LAC

+

GUS

+







LAC

+





+








1992)





non-salt adapted

E coli









TABLEOacute


K coli


Sampling

sites

AP

M l

M2

TR

PI

P2

N o of

strains

24

30

53

46

55

11

Thermo-

tolerant

No

24

28

51

42

51

9

100

93

96

91

93

82

Thermo-

sensitive

b

No

0

2

2

4

3

12

0

7

4

9

5

5

Indol-

No

0

2

3

2

1

9

0

7

6

4

2

4


k






REFERENCES


315(1996)



1-117


Bej


(1991)


i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5



1246 ALONSO ET AL


M icrobiol 59 3534-3544 (1993)




336-344 (1992)




Microbiol 57 1528-1534(1991)



J A W W A 84 98-104 (1992)





pp 48-58






(1994)






i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5




E COLI

1247


207-214 (1991)


2684 (1988)

Kampfer P

Rauhoff









foods




1476-1481 (1987)


59 786-790(1995)



M W at Sci T ec h 2 5 93-103 (1992)




Rice


(1990)







1248 ALONSO ET AL


(1993)


2236-2243 (1996)


(1976)



e c e i v e d

December

22 1997


i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5



24

ALONSO ET AL

TABLE 4

Num ber o f

coli

a nd N o n-E

coli



Reactions

Phenotype

LAC GUS

AP

C

M l

M 2

TR

PI

P2

Total

LAC

+

GUS

+

AP

M l

M 2

T R

PI

P2

Total

LACGUS

A P

M l

M 2

T R

P I

P2

Total

LAC GUS-

A P

M l

M2

TR

PI

P2

Total

Isolates

N o

37

31

42

41

31

11

193

24

29

48

48

52

11

212

2

9

0

3

2

0

16

17

23

15

36

46

29

166

E

N o

14

10

25

19

18

1

87

24

27

48

46

52

10

207

2

8

0

3

2

0

15

0

2

1

5

1

0

9

coli

( )

38

32

59

46

58

9

45

100

93

100

96

100

91

98

100

89

0

100

100

0

94

0

9

7

14

2

0

5

K E C

No

23

20

17

17

8

1

86

0

2

0

2

0

1

5

0

1

0

0

0

0

1

17

20

12

14

6

2

71

( )

62

68

41

42

26

9

45

0

7

0

4

0

9

2

0

11

0

0

0

0

6

100

87

80

39

13

7

43

Non

coliform

b

No

0

0

0

0

3

5

8

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

1

2

12

22

10

47

( )

0

0

0

0

10

46

4

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

4

13

33

48

34

28

N ot

identified

No

0

1

0

5

2

4

12

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

5

17

17

39

( )

0

3

0

12

6

36

6

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

14

37

59

24

KEC


b


Sampling sites



i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5



TAB LE 5


O

m

sect

o

L A C

+

G U S

N o

L A C

+


4

N o L A C G U S -

No


Klebsiella oxytoca

K Pneuntoniae

Citrobacterfreundii

C Amalonaticus

Escherichia coli

4

2

2

6

1

6

pound co

Cfreundii

8

1

ot l

E co

21

Pseudomonas

spp

P fluorescens

Acinetobacter

spp

Flavobacterium

spp

Proteus

spp

Salmonella typhi

Citrobacterfreundii

C amalonaticus

Klebsiella oxytoca

K pneumoniae


E agglomerans

E sakazakii

Escherichia coli

4

1

1

1

1

1

8

1

4

2

3

1

1

4

33

a

LA C

+


bull LAC GU S

+


l A C

G US

+


d





1242 ALONSO ET AL



1991










sample

itself


+

G U S

+










red colonies (LAC

+



7



i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5




G U S

+

colony and 5 LAC

+

GUS

+
























i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5



1244 ALONSO ET AL

When LAC

+

GUS LAC

+

GUS

+







LAC

+





+








1992)





non-salt adapted

E coli









TABLEOacute


K coli


Sampling

sites

AP

M l

M2

TR

PI

P2

N o of

strains

24

30

53

46

55

11

Thermo-

tolerant

No

24

28

51

42

51

9

100

93

96

91

93

82

Thermo-

sensitive

b

No

0

2

2

4

3

12

0

7

4

9

5

5

Indol-

No

0

2

3

2

1

9

0

7

6

4

2

4


k






REFERENCES


315(1996)



1-117


Bej


(1991)


i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5



1246 ALONSO ET AL


M icrobiol 59 3534-3544 (1993)




336-344 (1992)




Microbiol 57 1528-1534(1991)



J A W W A 84 98-104 (1992)





pp 48-58






(1994)






i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5




E COLI

1247


207-214 (1991)


2684 (1988)

Kampfer P

Rauhoff









foods




1476-1481 (1987)


59 786-790(1995)



M W at Sci T ec h 2 5 93-103 (1992)




Rice


(1990)







1248 ALONSO ET AL


(1993)


2236-2243 (1996)


(1976)



e c e i v e d

December

22 1997


i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5



TAB LE 5


O

m

sect

o

L A C

+

G U S

N o

L A C

+


4

N o L A C G U S -

No


Klebsiella oxytoca

K Pneuntoniae

Citrobacterfreundii

C Amalonaticus

Escherichia coli

4

2

2

6

1

6

pound co

Cfreundii

8

1

ot l

E co

21

Pseudomonas

spp

P fluorescens

Acinetobacter

spp

Flavobacterium

spp

Proteus

spp

Salmonella typhi

Citrobacterfreundii

C amalonaticus

Klebsiella oxytoca

K pneumoniae


E agglomerans

E sakazakii

Escherichia coli

4

1

1

1

1

1

8

1

4

2

3

1

1

4

33

a

LA C

+


bull LAC GU S

+


l A C

G US

+


d





1242 ALONSO ET AL



1991










sample

itself


+

G U S

+










red colonies (LAC

+



7



i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5




G U S

+

colony and 5 LAC

+

GUS

+
























i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5



1244 ALONSO ET AL

When LAC

+

GUS LAC

+

GUS

+







LAC

+





+








1992)





non-salt adapted

E coli









TABLEOacute


K coli


Sampling

sites

AP

M l

M2

TR

PI

P2

N o of

strains

24

30

53

46

55

11

Thermo-

tolerant

No

24

28

51

42

51

9

100

93

96

91

93

82

Thermo-

sensitive

b

No

0

2

2

4

3

12

0

7

4

9

5

5

Indol-

No

0

2

3

2

1

9

0

7

6

4

2

4


k






REFERENCES


315(1996)



1-117


Bej


(1991)


i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5



1246 ALONSO ET AL


M icrobiol 59 3534-3544 (1993)




336-344 (1992)




Microbiol 57 1528-1534(1991)



J A W W A 84 98-104 (1992)





pp 48-58






(1994)






i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5




E COLI

1247


207-214 (1991)


2684 (1988)

Kampfer P

Rauhoff









foods




1476-1481 (1987)


59 786-790(1995)



M W at Sci T ec h 2 5 93-103 (1992)




Rice


(1990)







1248 ALONSO ET AL


(1993)


2236-2243 (1996)


(1976)



e c e i v e d

December

22 1997


i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5



1242 ALONSO ET AL



1991










sample

itself


+

G U S

+










red colonies (LAC

+



7



i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5




G U S

+

colony and 5 LAC

+

GUS

+
























i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5



1244 ALONSO ET AL

When LAC

+

GUS LAC

+

GUS

+







LAC

+





+








1992)





non-salt adapted

E coli









TABLEOacute


K coli


Sampling

sites

AP

M l

M2

TR

PI

P2

N o of

strains

24

30

53

46

55

11

Thermo-

tolerant

No

24

28

51

42

51

9

100

93

96

91

93

82

Thermo-

sensitive

b

No

0

2

2

4

3

12

0

7

4

9

5

5

Indol-

No

0

2

3

2

1

9

0

7

6

4

2

4


k






REFERENCES


315(1996)



1-117


Bej


(1991)


i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5



1246 ALONSO ET AL


M icrobiol 59 3534-3544 (1993)




336-344 (1992)




Microbiol 57 1528-1534(1991)



J A W W A 84 98-104 (1992)





pp 48-58






(1994)






i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5




E COLI

1247


207-214 (1991)


2684 (1988)

Kampfer P

Rauhoff









foods




1476-1481 (1987)


59 786-790(1995)



M W at Sci T ec h 2 5 93-103 (1992)




Rice


(1990)







1248 ALONSO ET AL


(1993)


2236-2243 (1996)


(1976)



e c e i v e d

December

22 1997


i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5




G U S

+

colony and 5 LAC

+

GUS

+
























i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5



1244 ALONSO ET AL

When LAC

+

GUS LAC

+

GUS

+







LAC

+





+








1992)





non-salt adapted

E coli









TABLEOacute


K coli


Sampling

sites

AP

M l

M2

TR

PI

P2

N o of

strains

24

30

53

46

55

11

Thermo-

tolerant

No

24

28

51

42

51

9

100

93

96

91

93

82

Thermo-

sensitive

b

No

0

2

2

4

3

12

0

7

4

9

5

5

Indol-

No

0

2

3

2

1

9

0

7

6

4

2

4


k






REFERENCES


315(1996)



1-117


Bej


(1991)


i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5



1246 ALONSO ET AL


M icrobiol 59 3534-3544 (1993)




336-344 (1992)




Microbiol 57 1528-1534(1991)



J A W W A 84 98-104 (1992)





pp 48-58






(1994)






i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5




E COLI

1247


207-214 (1991)


2684 (1988)

Kampfer P

Rauhoff









foods




1476-1481 (1987)


59 786-790(1995)



M W at Sci T ec h 2 5 93-103 (1992)




Rice


(1990)







1248 ALONSO ET AL


(1993)


2236-2243 (1996)


(1976)



e c e i v e d

December

22 1997


i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5



1244 ALONSO ET AL

When LAC

+

GUS LAC

+

GUS

+







LAC

+





+








1992)





non-salt adapted

E coli









TABLEOacute


K coli


Sampling

sites

AP

M l

M2

TR

PI

P2

N o of

strains

24

30

53

46

55

11

Thermo-

tolerant

No

24

28

51

42

51

9

100

93

96

91

93

82

Thermo-

sensitive

b

No

0

2

2

4

3

12

0

7

4

9

5

5

Indol-

No

0

2

3

2

1

9

0

7

6

4

2

4


k






REFERENCES


315(1996)



1-117


Bej


(1991)


i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5



1246 ALONSO ET AL


M icrobiol 59 3534-3544 (1993)




336-344 (1992)




Microbiol 57 1528-1534(1991)



J A W W A 84 98-104 (1992)





pp 48-58






(1994)






i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5




E COLI

1247


207-214 (1991)


2684 (1988)

Kampfer P

Rauhoff









foods




1476-1481 (1987)


59 786-790(1995)



M W at Sci T ec h 2 5 93-103 (1992)




Rice


(1990)







1248 ALONSO ET AL


(1993)


2236-2243 (1996)


(1976)



e c e i v e d

December

22 1997


i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5




TABLEOacute


K coli


Sampling

sites

AP

M l

M2

TR

PI

P2

N o of

strains

24

30

53

46

55

11

Thermo-

tolerant

No

24

28

51

42

51

9

100

93

96

91

93

82

Thermo-

sensitive

b

No

0

2

2

4

3

12

0

7

4

9

5

5

Indol-

No

0

2

3

2

1

9

0

7

6

4

2

4


k






REFERENCES


315(1996)



1-117


Bej


(1991)


i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5



1246 ALONSO ET AL


M icrobiol 59 3534-3544 (1993)




336-344 (1992)




Microbiol 57 1528-1534(1991)



J A W W A 84 98-104 (1992)





pp 48-58






(1994)






i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5




E COLI

1247


207-214 (1991)


2684 (1988)

Kampfer P

Rauhoff









foods




1476-1481 (1987)


59 786-790(1995)



M W at Sci T ec h 2 5 93-103 (1992)




Rice


(1990)







1248 ALONSO ET AL


(1993)


2236-2243 (1996)


(1976)



e c e i v e d

December

22 1997


i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5



1246 ALONSO ET AL


M icrobiol 59 3534-3544 (1993)




336-344 (1992)




Microbiol 57 1528-1534(1991)



J A W W A 84 98-104 (1992)





pp 48-58






(1994)






i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5




E COLI

1247


207-214 (1991)


2684 (1988)

Kampfer P

Rauhoff









foods




1476-1481 (1987)


59 786-790(1995)



M W at Sci T ec h 2 5 93-103 (1992)




Rice


(1990)







1248 ALONSO ET AL


(1993)


2236-2243 (1996)


(1976)



e c e i v e d

December

22 1997


i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5




E COLI

1247


207-214 (1991)


2684 (1988)

Kampfer P

Rauhoff









foods




1476-1481 (1987)


59 786-790(1995)



M W at Sci T ec h 2 5 93-103 (1992)




Rice


(1990)







1248 ALONSO ET AL


(1993)


2236-2243 (1996)


(1976)



e c e i v e d

December

22 1997


i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5



1248 ALONSO ET AL


(1993)


2236-2243 (1996)


(1976)



e c e i v e d

December

22 1997


i t y ] a t 2 1 0 5 1 8 J u l y 2 0

1 5

alonso, 1998. quantitative determination of e. coli and fecal coliforms in water using a chromogenic...

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