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Vesterlund Et Al, 2007 - Safety Assessment of Lactobacillus Strains PDF
Vesterlund Et Al, 2007 - Safety Assessment of Lactobacillus Strains PDF
www.elsevier.com/locate/ijfoodmicro
Department of Biochemistry and Food Chemistry and Functional Foods Forum, University of Turku, 20014 Turku, Finland
b
Laboratory of Medical Microbiology, University of Antwerp, 2610 Wilrijk, Belgium
c
Valio Ltd, R&D, 00039 Helsinki, Finland
d
Department of Medical Microbiology, Leiden University Medical Center, 2300 RC Leiden, The Netherlands
Received 13 October 2006; received in revised form 10 February 2007; accepted 14 February 2007
Abstract
The widespread use of immunosuppressive therapy and antimicrobial agents as well as novel probiotics without a long history of safe use has
increased requirements for safety assessment of probiotic lactobacilli. In this study 44 faecal, 52 blood and 15 probiotic isolates (including 3 dairy
strains) of Lactobacillus were assayed for their adhesion properties to extracellular matrix proteins and mucus, hemolysis, ability to avoid the
induction of respiratory burst in peripheral blood mononucleocytes (PMN) and resistance to human serum. Among tested strains adhesion to
collagen, fibrinogen and mucus was isolate-specific and no statistically significant differences were obtained between faecal, blood and probiotic
isolates. However, blood isolates showed a trend for higher adhesion to mucus than probiotic strains (P = 0.07). Probiotic strains induced lower
respiratory burst in PMN when compared to the blood isolates (P b 0.05). Moreover, there was a positive correlation between adhesion to collagen
and induction of respiratory burst for faecal isolates (P b 0.05). In the determination of serum resistance, probiotic strains showed a trend for lower
sensitivity to human serum-mediated killing when compared to the faecal isolates (P = 0.07). None of the measurable virulence factors were found
to be present at statistically higher level in clinical blood isolates when compared to faecal and/or probiotic isolates indicating that these factors do
not cause risk when safety of probiotics is considered. However, the significance of adhesion to mucus, low induction of respiratory burst in PMN
and resistance to human serum-mediated killing may need further evaluation in experimental animal models and in epidemiological data.
2007 Elsevier B.V. All rights reserved.
Keywords: Probiotic; Lactobacillus; Safety; Risk factor
1. Introduction
Lactobacilli are generally regarded as safe due to their long
history of safe use in fermented foods and their presence in the
normal intestinal and urogenital microbiota of humans (Ishibashi and Yamazaki, 2001; Tuohy et al., 2003). However,
lactobacilli have been associated with isolated cases of
infections (Cannon et al., 2005; De Groote et al., 2005; Farina
et al., 2001; Horwitch et al., 1995; Husni et al., 1997; Land
et al., 2005; Mackay et al., 1999; Rautio et al., 1999; Salminen
Corresponding author. Tel.: +358 2 3336861; fax: +358 2 3336862.
E-mail address: satu.vesterlund@utu.fi (S. Vesterlund).
1
Present address: Danisco Innovation, 02460 Kantvik, Finland.
0168-1605/$ - see front matter 2007 Elsevier B.V. All rights reserved.
doi:10.1016/j.ijfoodmicro.2007.02.002
326
Table 1
Number of isolates used in the study per Lactobacillus species
Species
Faecal isolates
Probiotic strains
L. acidophilus
L. amylovorus
L. animalis
L. casei
L. coprophilus
L. crispatus
L. curvatus
L. delbrueckii
L. fermentum
L. gasseri
L. jensenii
L. johnsonii
L. paracasei
L. plantarum
L. reuteri
L. rhamnosus
L. sakei
L. salivarius
L. zeae
Total number of
isolates
3
1
1
2
1
1
1
3
1
7
3
2
17
44
4
2
2
1
5
2
1
25
2
1
1
52
1 (dairy)
2
2
1 (dairy)
2
3 (1 dairy)
15
For statistical comparisons three dairy strains were included into probiotics
group.
327
2.3. Hemolysis
Hemolysis was tested by the modified method of Baumgartner et al. (1998). Strains were grown on MRS agar supplemented
with 5% human blood (group O) and incubated for 48 h at 37 C
under anaerobic conditions. Bacillus cereus, grown under
aerobic conditions, was included as a positive control.
Fig. 1. Adhesion of faecal, clinical blood and probiotic Lactobacillus isolates to collagen (A), fibrinogen (B) and mucus (C). Lines indicate median, boxes 25th and
75th percentiles, whiskers indicate 5th and 95th percentiles.
328
Fig. 3. Survival in human serum of faecal, clinical blood and probiotic Lactobacillus isolates. Represented as percentage survival when survival in human
serum is compared to survival in heat-inactivated human serum. Lines indicate
median, boxes 25th and 75th percentiles, whiskers indicate 5th and 95th
percentiles.
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