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Molecular Diversity and Industrial<br />
Characteristics of Streptococcus<br />
Streptococcus<br />
thermophilus<br />
th th thermophilus hil hil<br />
Walid Walid MM. El El-Sharoud Sharoud, PhD PhD<br />
Food Safety and Microbial Physiology Laboratory<br />
Faculty of Agriculture<br />
Mansoura University
Streptococcus Streptococcus thermophilus<br />
thermophilus<br />
-A Major j Industrial Bacterium<br />
that is massively used in the<br />
preparation of dairy products. products<br />
-Its Its Average Annual Market is<br />
around aroundd 40 billi billion USD) USD).
Safety of S. S. thermophilus<br />
thermophilus<br />
- Generally Regarded as Safe (GRAS) and and has<br />
has<br />
been granted the “Qualified Presumption of<br />
Safety” Safety Safety” (QPS) Status Status in in Europe.<br />
Europe
SS.<br />
. thermophilus<br />
thermophilus is<br />
An ANGEL among the DEVILS!!<br />
S. pyogenes<br />
S. pneumoniae p SS. mutans mutans<br />
S. thermophilus<br />
S. vestibularis
How Could SS. . thermophilus<br />
thermophilus Manage to<br />
Become A Safe Bacterium?!!<br />
C Complete l t G Genome S Sequence and d C Comparative ti<br />
Analysis of SS.. thermophilu thermophilus suggested that that:<br />
- Virulence genes found in other pathogenic streptoocci<br />
have been inactivated or deleted in SS.. thermophilus<br />
thermophilus..<br />
-This This was the result of numerous Horizontal Gene Transfer<br />
events between SS.. SS.. thermophilus<br />
thermophilus thermophilus<br />
thermophilus and other milk<br />
fermenting Lactic acid bacteria such as Lactobacillus<br />
Lactobacillus<br />
bulgaricus, bulgaricus, Lactococcus Lactococcus lactis, lactis, etc.. etc..<br />
- Ecological Proximity has been thus more important than<br />
Ph Phylogenetic l ti P Proximity i it i in Dt Determining ii th the Ft Fate of f SS SS..<br />
thermophilus.<br />
thermophilus
Research Objectives<br />
Objectives<br />
- To study the genetic diversity of SS SS.. thermophilus<br />
thermophilus<br />
thermophilus<br />
thermophilus<br />
strains isolated from traditional dairy products. products<br />
-To To examine the technological characteristics of SS..<br />
thermophilus<br />
thermophilus thermophilus<br />
thermophilus isolated from traditional dairy<br />
products products.
Isolation of S. S. thermophilus<br />
thermophilus from Traditional<br />
Egyptian Dairy Dairy Products<br />
Products<br />
Collection of of 85 samples of Kariesh cheese<br />
cheese<br />
and Laban Rayeb from the Mansoura City during 2008-2009 2008 2009<br />
Gram Staining<br />
Sample Preparation<br />
Plating on Elliker Agar<br />
Morphological and Physiological Examinations of<br />
Suspect S. S. thermophilus<br />
thermophilus Colonies<br />
Catalase test<br />
Growth at 10 10ºC ºC & 45 45ºC ºC<br />
Growth in 4 & 6.5% % NaCl
serB serB<br />
PCR Identification of Promising SS. SS .<br />
thermophilus<br />
thermophilus
AT A Total t l of ff70<br />
70 SS. SS . thermophilus<br />
th th thermophilus hil hil<br />
Strains Could be Confirmed<br />
SSamples l NNo. of fconfirmed fi dSS.<br />
thermophilu strains<br />
Kariesh Cheese<br />
56<br />
Laben Rayeb<br />
Total<br />
14<br />
70
11- PCR<br />
Multilocus Sequence Sequence Typing Typing (MLST) of<br />
of<br />
Confirmed SS. . thermophilus<br />
thermophilus<br />
Strains Targeting 6 Housekeeping Housekeeping Genes<br />
22- DNA Seq Sequencing encing of<br />
of<br />
PCR products.<br />
3- Database & Software<br />
Analyses.
MLST R Results lt of f SS. SS . thermophilus<br />
th th thermophilus hil hil<br />
Strains<br />
Strain Source ST glcK ddlA tkt proA ptsI serB<br />
JIM JIM9139 9139 Kariesh 94 17 2 15 5 15 22<br />
JIM JIM10172 10172 Kariesh 106 6 2 15 5 15 13<br />
JIM JIM101 JIM JIM10171 10171 101 1 K Kariesh i h 106 6 2 115 5 15 13<br />
JIM JIM9151 9151 Kariesh 106 6 2 15 5 15 13<br />
JIM JIM10169 10169 Laben Rayeb 106 6 2 15 5 15 13<br />
JIM JIM10170 JIM JIM10170 10170 Laben Rayeb Rayeb 106 6 2 15 5 15 13<br />
JIM JIM10173 10173 Laben Rayeb 107 6 2 15 2 6 13<br />
JIM JIM10174 10174 Kariesh 108 6 2 18 5 15 13<br />
JIM JIM9150 9150 Laben Rayeb 108 6 2 18 5 15 13<br />
JIM JIM9160 9160 Laben Rayeb 109 6 2 18 5 15 26<br />
JIM JIM10175 10175 Kariesh 110 1 2 2 3 25 3<br />
JIM JIM10176 10176 Kariesh 111 1 1 17 23 3 23<br />
JIM JIM9156 9156 Kariesh 111 1 1 17 23 3 23<br />
JIM JIM10177 10177 Kariesh 112 19 2 16 23 3 23<br />
JIM JIM10178 10178 Kariesh 113 19 2 16 23 3 24<br />
JIM JIM9154 JIM JIM9154 9154 Kariesh 114 19 1 16 2 3 24<br />
JIM JIM10179 10179 Kariesh 115 7 2 4 2 22 10<br />
JIM JIM10180 10180 Laben Rayeb 116 6 2 15 5 15 25
0,001<br />
JIM7446 Levain<br />
JIM10180-Egypt-2009<br />
JIM10177-Egypt-2009<br />
JiM9150-Egypt-2009<br />
JIM10174-Egypt-2009<br />
JIM10172-Egypt-2008<br />
JiM9151-Egypt-2009<br />
JiM9151 Egypt 2009<br />
JIM10171-Egypt-2009<br />
JIM10170-Egypt-2009<br />
JIM10169-Egypt-2009<br />
JIM10173-Egypt-2009<br />
JIM10119 Inde-fromage traditionnel-2004<br />
JIM9139 Yaourt Egypt-2008<br />
JIM10178-Egypt-2009<br />
JiM9154-Egypt-2009<br />
CNRZ1595 Autriche-Emmental-1979<br />
CNRZ778-levain lactique a yaourt-Bulgari<br />
JIM10155 lait de traite-2009 (C5)<br />
JIM1004-Bulgaria-environment<br />
g<br />
CNRZ759 Bulgarie-LevainCY-1978<br />
LMG18311 UK-Yaourt-1974<br />
CNRZ385 Japon-Yaourt sucre-1971<br />
CNRZ406 levain lactique a yaourt - 1971<br />
CNRZ389 Levain yaourt-1971<br />
CNRZ403 CNRZ403-levain levain lactique a yaourt yaourt-1971 1971<br />
CNRZ446 Ivry sur seine-Yaourt-1973<br />
CNRZ386-levain lactique a yaourt-1971<br />
JIM10093 Haute Savoie-Lactoserum-1984<br />
CNRZ388 LevainCY-1971<br />
JIM7455 Levain<br />
CNRZ1066 LevainCY-1986<br />
CNRZ404-levain lactique a yaourt-1971<br />
CNRZ1588 Italie-fromage- 1962<br />
CNRZ160 levain lactique commercial a yao<br />
CNRZ779 levain lactique Boll 1979<br />
CNRZ1447 IInde d DDahli-Lait hli L i ffermente-1990 1990<br />
JiM9160-Egypt-2009<br />
A Phylogenetic tree based on concatenated sequences of 6 housekeeping<br />
genes in SS. . thermophilus<br />
thermophilus strains
CNRZ1447 Inde Dahli-Lait fermen mente-1990<br />
0.001<br />
JiM9160-Egypt-2 pt-2009<br />
JIM10119 Indee-fromage f traditionnel-2004<br />
JIM9139 39 Yaourt Egypt-2008<br />
JI JIM10173-Egypt-2009<br />
JIM10169-Egypt-2009<br />
JIM10170-Egypt-2009<br />
JIM10171-Egypt-2009<br />
JiM9151-Egypt-2009<br />
JIM10172-Egypt-2008<br />
JIM10174-Egypt-2009<br />
JiM9150-Egypt-2009<br />
JIM10093 Hau ute Savoie-Lactoserum-19 984<br />
CNRZ386 86-levain lactique a yaourt-1 -1971<br />
CNRZ44 446 Ivry sur seine-Yaourt-19 973<br />
CNRZ4 Z403-levain lactique a yaour urt-1971<br />
CNRZ RZ406 levain lactique a yaour urt - 1971<br />
CCNRZ385<br />
Japon-Yaourt suc ucre-1971<br />
CNRZ759 Bulgarie-LevainC nCY-1978<br />
CNRZ778-levain lactique a ya yaourt-Bulgari<br />
JIM10155 lait de traite-2 e-2009 (C5)<br />
JIM1004-Bulgaria-envi nvironment<br />
JIM10178 78-Egypt-2009<br />
JiM91 9154-Egypt-2009<br />
JIM10 M10177-Egypt-2009<br />
CNRZ160 levain lactiq que commercial a yao<br />
CNRZ404-levai ain lactique a yaourt-1971<br />
CNRZ779 leva vain lactique Boll 1979<br />
CNRZ15 588 Italie-fromage- 1962<br />
CNNRZ1066<br />
LevainCY-1986 86 1<br />
2<br />
CNRZ388 LevainCY-19 971<br />
CNRZ389 Levain yaourt urt-1971<br />
LMG18311 UK-Yao aourt-1974<br />
JIM7455 Levain n<br />
CNRZ1595 Aut Autriche-Emmental-1979<br />
JIM10180-Egypt-2009<br />
CNRZ27-bioghurt-1960 1960<br />
JIM10098 fromage frai<br />
JIM10106 Levain vain naturel naturel-1999 19<br />
JIM10089 89 Haute Haute Savoie-Lactoserum Savoie-Lactoserum-1985 1985<br />
198<br />
CNR NRZ1202 Grece Heraclion-Yaourt grec-198 198<br />
CNRZ1 RZ1213 Grece Heraclion-Yaourt grec-198 98<br />
JIM7446 Levain<br />
e frais-1999<br />
JIM10059 M10059 Savoie Savoie-Lactoserum Lactoserum-1986 1986<br />
JIM10032 Haute Savoie-Lait-1999<br />
Savoie-Lait<br />
JIM10024 Haute Savoie-Pate P te Pre Pressee Cuite<br />
CNRZ1157-Grece Heraclion-Yaourt grec-19<br />
C<br />
JIM1008 08-Bulgaria-environment<br />
CNRZ702 702/JIM8392 Grece Athenes-Yaourt gre gre<br />
JIM1005- 05-Bulgaria-environment<br />
JIM1007-Bulg ulgaria-environment<br />
CNRZ785-lev evain lactique-1979<br />
JIM1001-Bulg lgaria-environment-1997<br />
JIM1002-Bulgari ria-environment-1997<br />
CNRZ758 Bulgarie ie-LevainCY-1978<br />
CNRZ447-Netherland d-Yaourt 1973<br />
- 197<br />
JIM10124 pate ate molle-2006<br />
JIM1006-Bulgar aria-environment<br />
JIM10014-Bulga aria-environment<br />
JIM1003-Bulgari ria-environment<br />
CNRZ308-levain artisa sanal a gruyere de co<br />
CNRZ761-levain lactique ue a yaourt-1978<br />
JIM10117 J Inde-fromage trad aditionnel-2004<br />
ge frais-1999<br />
CNRZ1237<br />
CNRZ1593<br />
JIM10095 fromage frais<br />
CNRZ1156-Crete-Yaourt grec-1987<br />
CNRZ1208-Crete-Yaourt grec-1987<br />
1970<br />
CNRZ781 19979<br />
JIM10013-Bulga aria-environment<br />
JIM1009-Bulgaria-en nvironment-1998<br />
CNRZ26-levain C artisanal agruyere a g de com<br />
JIM1<br />
JIM10011-Bulgaria-e environment<br />
M10010-Bulgaria-environm ment-1998<br />
JIM10116 Slovaq quie-fromage traditionnel<br />
CNRZ368 CN NL-LevainCY-1<br />
TINA Sammito 38 38<br />
TINA Sammito 56 56<br />
CNRZ409 leva vain lactique commercial<br />
ere<br />
TINA Sammito32<br />
Com<br />
CNRZ456 levain n lactique a gruyere de C<br />
CNRZ21 Lemuy- y-levain artisanal a gruyer<br />
TINA Sammito 44<br />
TINA Lissandrello 19<br />
CNRZ24B Jura-L -Levain Comte-1959<br />
CNRZ22-levain artisana nal a gruyere de com<br />
CNRZ1120 Leva vain Comte-1987<br />
CNRZ1579-Itali ia-1962<br />
TTINA<br />
Labichino 110<br />
TINA Spata 66<br />
JIM10156 Fro omage -2009 (C5)<br />
urel-200<br />
Haute Savoie-Levain natu<br />
JIM10026 Haute H Savoie-Pate Press see Cuite<br />
JIM10114 4Levain naturel-1999<br />
JIM10037 Ha<br />
Haute Savoie-Levain natu<br />
JIM10042 H<br />
JIM10045<br />
CNRZ1575 75Italie-frompm-1962<br />
urel-200<br />
JIM10033 Haute H Savoie-Levain natu urel-200<br />
aturel-200<br />
5Haute Savoie-Levain na<br />
JIM10039 Haute H Savoie-Levain nat aturel-200<br />
JIM10050 Haute Ha Savoie-Levain nature rel-200<br />
JIM10054 Haute Ha Savoie-Levain natu urel-200<br />
JIM10034 Haute Ha Savoie-Levain natu urel-200<br />
JIM10022 Haute Ha Savoie-Pate Presse see Cuite<br />
JIM10076 76 Haute Savoie-Lactoserum um-1985<br />
JIM10104 Levain L naturel-1999<br />
JIM10099 Lait L cru-2001<br />
JIM10112 Levain Le naturel-1999<br />
JIM10100 00 Lait cru-2001<br />
JIM10014 Haute Savoie-Pate S Pressee Cuite<br />
JIM10047 Haute Savoie-Levain Sa naturel-200<br />
JIM10085 Haute Savoie-Lactoserum-1986<br />
S<br />
JIM10055 Isere-La Lactoserum-1985<br />
JiM9156-Egypt-2009<br />
JIM10176-Egypt-2008 8<br />
JIM10012-Bulgaria-environment<br />
CNRZ457-levain artisanal aa ggruyere<br />
de co<br />
6<br />
JIM10120 pate molle-2006 006<br />
JIM10123 pate molle-2006<br />
JIM 10079 Haute Savoie ie-Lactoserum-1984<br />
JIM 10066 Savoie-Lacto ctoserum-1985<br />
JIM 10064Savoie-Lacto ctoserum-1984<br />
JIM 10060 Savoie-Lac Lactoserum-1985<br />
JIM 10020 Haute Sav avoie-Pate Pressee Cuit<br />
JIM10021 Haute Savoie-Pate Pressee Cuite<br />
JIM10013 Haute Savoie-Pate Pressee Cuite<br />
JIM10121 fromage traditionnel-2004<br />
JIM 10001 Haute Savoie-Pate Pressee Cuit<br />
JIM10151 sol-2009 (C4)<br />
JIM10153 Environnement-2009 (C5)<br />
JIM10359 fromagerie-2008 (C3)<br />
CNRZ1584-Italia-1962<br />
CNRZ1587<br />
JIM10175 JIM10175-Egypt-2009<br />
Egypt 2009<br />
JIM10254 JIM10254 trayon t Toupie (C1)<br />
LMD9 LMD9 UUSA-fromage<br />
CNR CNRZ310 Barretaine-levain artisanal a gr<br />
CNR CNRZ302-levain artisanal a gruyere de co<br />
CN CNRZ1592 Allemagne-LevainCY-1987<br />
CN CNRZ703 Mongolie-Lait fermente-1974<br />
CN CNRZ701Finlande Helsinki-levain a emment<br />
JIM JIM10300 Fromage J+2 2nde Campagne (JIM (JIM1<br />
JI JIM 7444 Levain<br />
CCH CCHSS9 Hospital-2002<br />
JJIM10087<br />
Haute Savoie-Lactoser erum-1984<br />
JIM 7442 Levain -<br />
JIM 7443 Levain -<br />
JIM10118 ferment-2000<br />
CNRZ1691 Irelande-LevainCY-1992<br />
JIM10139 ferment yaourt rt -2008 (C2)<br />
JIM 10006 Haute Savoi oie-Pate Pressee Cuit<br />
JIM10031 Haute Savoie-Lait-19 1999<br />
JIM8232 Lait-2002<br />
JIM10097 fromage frai ais-1999<br />
JIM10058 Haute Sao one-Lactoserum-1988<br />
JIM10078 Haute Sav avoie-Lactoserum-1986<br />
JIM10016 Haute SSavoie-Pate<br />
Pressee Cuit te<br />
JIM10018 Haute SSavoie-Pate<br />
Pressee Cuit uite<br />
JIM10052Haute SSavoie-Levain<br />
naturel-200 001<br />
JIM10256 Fe Ferment yaourt -2007 (C1) )<br />
JIM10096 96 fromage frais-1999<br />
JIM10179 79-Egypt-2008<br />
TINA Spa pata 86<br />
TINA Sppata<br />
64<br />
TINA A Spata 71<br />
JIM103 0358 lait empresurage-200 008 (C3)<br />
JIMM10152<br />
lait traite-fromage ge-2009 (C4)<br />
JIMM10154<br />
lait de traite +200 009 (C5)<br />
TINA Spata 88<br />
TINA Spata 93<br />
JIM10010 Haute ute Savoie-Pate Pressee CCuite<br />
JIM10072 Hau aute Savoie-Lactoserum-19 1986
I Industrial d t i l Traits T it of f SS. SS . thermophilus<br />
th th thermophilus hil hil<br />
Strains<br />
Test<br />
No of positive strains<br />
(%)<br />
Lactose Galactose Urease Exopolysaccharide<br />
fermentation fermentation Production Production<br />
70 (100%) 27<br />
(40%)<br />
5<br />
(10%)<br />
3<br />
(5%)<br />
No of Negative 0 43 65 67<br />
strains (%) (0%) (60%) (90%) (95%)<br />
Total 70 70 70 70
Conclusions<br />
- A discrete level of genetic and phenotypic<br />
diversity does exist among SS.. thermophilus<br />
thermophilus<br />
isolated from traditional Egyptian dairy<br />
products products.
Acknowledgments<br />
- This work has been carried out in collaboration between the FSMP<br />
lab at Mansoura University y and the French “Laboratoire de Génétique q<br />
Microbienne” lab at INRA within the context of the Egypt-France<br />
Egypt France<br />
Science and Technology Cooperation Programme (IMHOTEP).
Thanks
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