Antimicrobial Activity of Novel Probiotic Strains against ... · bactericidal activity against...

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Antimicrobial Activity of Probiotic Strains by Well Diffusion Agar Method

Antimicrobial Susceptibility Profile

The antimicrobial activity of probiotic strains tested by

broth assay showed inhibition of Listeria monocytogenes

up to 6.60 log10 CFU/ml and probiotic growth of up to 8.66

log10 CFU/ml .

Probiotic strains analyzed in this study were able to

reduce Salmonella, E. coli O157:H7, and Listeria

monocytogenes.

A similar effect was observed when comparing the

antimicrobial effect of overnight cultures to cell-free

suspensions.

Probiotics showed bacteriostatic activity against

Salmonella and E. coli O157:H7, compared to a

bactericidal activity against Listeria monocytogenes.

From the 28 selected strains, 3 were selected for

further study to reduce Listeria monocytogenes in

ready-to-eat meats.

Figure 9: Inhibition

of Listeria

monocytogenes by

probiotic J19 on

timepoint 6 hours

Figure 8: Inhibition of Listeria

monocytogenes by probiotic

J19, J7, J34, J27, J25, J43 on

timepoint 6 hours

Figure 7: Inhibition

of Listeria

monocytogenes by

probiotic L25 on

timepoint 6 hours

Antimicrobial Activity of Probiotic Strains by Broth assay

From the selected panel of 28 probiotic strains, 13 probiotic strains had antimicrobial activity on the agar

well diffusion assay against Salmonella with a maximum inhibition zone of 12 mm; 13 strains for E. coli

O157:H7 with 9mm; and all 28 strains showed antimicrobial activity against L. monocytogenes with up

to 26 mm of inhibition.

Figure 2: Inhibition of pathogens by probiotic strains

Listeria monocytogenes

E.Coli O157:H7

Salmonella

Antimicrobial Activity of Novel Probiotic Strains against Listeria

monocytogenes, Salmonella and E. coli O157:H7Valencia, María Jose1; Ayala, Diana,PhD2; Franco, Jorge2; Brashears, Mindy,PhD2

1 SOWER Scholar, 2 Texas Tech University

13 13

28

In the United States, 9.4 million episodes of foodborne

illness are caused by 31 major pathogens, from which Listeria

monocytogenes, Salmonella and E. coli O157:H7 are among

the most common causes of foodborne infections. Indeed,

over 40,000 and 3,500 laboratory-confirmed Salmonella and

E. coli O157:H7 cases respectively are reported annually to

the Centers for Disease Control and Prevention. On the other

hand, an estimated 1,600 people get listeriosis each year, and

about 260 die. Probiotics are a natural alternative to

counteract foodborne pathogens, they are live

microorganisms that when administered in adequate amounts

confer beneficial health effects to the host. Lactic acid bacteria

are the most commonly used group of bacteria as probiotic

strains, they produce antimicrobial compounds such as

bacteriocins able to reduce and/or inhibit human foodborne

pathogens. This study aimed to characterize potential

probiotic strains to reduce Salmonella, E. coli O157:H7, and

Listeria monocytogenes.

Abstract Results

A natural alternative to counteract foodborne pathogens are

probiotics, live microorganisms that when administered in

adequate amounts confer beneficial health effects to the host.

Probiotics are also an alternative to decrease the use of

antibiotics in animal feed, which could have potential risks

such as the development of antimicrobial resistance and the

spread of antibiotic resistance genes.

Results

Figure 3: Inhibition of Listeria monocytogenes by

probiotic J27 with a zone of inhibition of 21.5 mm, and

probiotic J7 with 25.5 mm.

Introduction

Methods

5.014

9.136 8.778 8.204

4.523 2.921 2.477 2.731

0.000

2.000

4.000

6.000

8.000

10.000

0 hrs 6 hrs 12 hrs 24 hrs

Log

CFU

/cm

2

HoursFigure 4: Probiotic J14

Probiotic Listeria m.

5.286

9.186 8.637 8.067

4.523 3.125 2.564 2.810

0.000

2.000

4.000

6.000

8.000

10.000

0 hrs 6 hrs 12 hrs 24 hrs

Log

CFU

/cm

2

HoursFigure 6: Probiotic J16

Probiotic Listeria m.

5.824

9.176 8.778 8.669

4.5233.308 2.778 2.354

0.000

2.000

4.000

6.000

8.000

10.000

0 hrs 6 hrs 12 hrs 24 hrs

Log

CFU

/cm

2

HoursFigure 5: Probiotic L20-B

Probiotic Listeria m.

10.5 11 11.514

11.5 10.5 1114

9 914

9

18

10.5 9

25.5 25 25

10 9

2623

20.524 24.5 25 25.5

21.5

8 9 6.5

8

7 7 6

8

7

0

0

0

0

77

7 7 7

0 0

0

00

0 0 0 0

078

7

6

6 6 7

8

7

7

6

6

12

0

0 0 0

0 0

0

00

0 0 0 0

0

0

5

10

15

20

25

30

35

L2-A L3-A L4-A L4-B L5-B L22 L23-A L28 L25 L1 L19 L23-B L24-B L8-B L21 J7 J34 J43 L11 L12 J19 L3-A L8-A L20-B J14 J16 J25 J27In

hib

itio

n z

on

e (

mm

)Probiotics

Figure 1: Well Diffusion Agar method

Listeria monocytogenes (mm) E. coli O157:H7 (mm) Salmonella (mm)

200 strains of probiotics isolated from meat and vegetables samples

Well diffusion method

Broth assayAntimicrobial

resistance

28 most effective strains selected

0 2 4 6 8 10 12 14 16 18

Ampicilin

Clindamycin

Daptomycin

Erythromycin

Gentamicin

Levofloxacin

Linezolid

Penicillin

Rifampicin

Synercid

Tetracycline

Tmp/Sxt

Vancomycin

No.IsolatesResistant

No.IsolatesSusceptible

ResultsConclusion

Figure 10: Antimicrobial

resistance