Food Safety and Microbial Physiology Laboratory. Mansoura University
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1 Molecular Diversity and Industrial Characteristics of Streptococcus thermophilus hil Walid M. El-Sharoud, PhD Food Safety and Microbial Physiology Laboratory Faculty of Agriculture Mansoura University
2 Streptococcus thermophilus -A Major Industrial Bacterium that is massively used in the preparation of dairy products. -Its Average Annual Market is around 40 billion USD).
3 Safety of S. thermophilus - Generally Regarded as Safe (GRAS) and has been granted the Qualified Presumption of Safety (QPS) Status in Europe.
4 S. thermophilus is An ANGEL among the DEVILS!! S. pyogenes S. pneumoniae S. mutans S. thermophilus S. vestibularis
5 How Could S. thermophilus Manage to Become A Safe Bacterium?!! Complete Genome Sequence and Comparative Analysis of S. thermophilus suggested that: - Virulence genes found in other pathogenic streptoocci have been inactivated or deleted in S. thermophilus. -This was the result of numerous Horizontal Gene Transfer events between S. thermophilus and other milk- fermenting Lactic acid bacteria such as Lactobacillus bulgaricus, Lactococcus lactis, etc.. - Ecological Proximity has been thus more important than Phylogenetic Proximity it in Dt Determining ii the Ft Fate of S. thermophilus.
6 Research Objectives - To study the genetic diversity of S. thermophilus strains isolated from traditional dairy products. -To examine the technological characteristics of S. thermophilus isolated from traditional dairy products.
7 Isolation of S. thermophilus from Traditional Egyptian Dairy Products Collection of 85 samples of Kariesh cheese and Laban Rayeb from the Mansoura City during Sample Preparation Plating on Elliker Agar Morphological and Physiological Examinations of Suspect S. thermophilus Colonies Gram Staining Catalase test Growth at 10ºC & 45ºC Growth in 4 & 6.5% NaCl
8 PCR Identification of Promising S. S thermophilus serb
9 AT Total of f70 S. S thermophilus hil Strains Could be Confirmed Samples No. of confirmed ds. thermophilus strains Kariesh Cheese 56 Laben Rayeb Total 14 70
10 Multilocus Sequence Typing (MLST) of Confirmed S. thermophilus Strains Targeting 6 Housekeeping Genes 1- PCR 2- DNA Sequencing encing of PCR products. 3- Database & Software Analyses.
11 Strain MLST Results of S. S thermophilus hil Strains Source ST glck ddla JIM9139 Kariesh JIM10172 Kariesh JIM Kariesh JIM9151 Kariesh JIM10169 Laben Rayeb JIM10170 Laben Rayeb JIM10173 Laben Rayeb JIM10174 Kariesh JIM9150 Laben Rayeb JIM9160 Laben Rayeb JIM10175 Kariesh JIM10176 Kariesh JIM9156 Kariesh JIM10177 Kariesh JIM10178 Kariesh JIM9154 Kariesh JIM10179 Kariesh JIM10180 Laben Rayeb tkt proa ptsi serb
12 JIM7446 Levain JIM10180-Egypt-2009 JIM10177-Egypt-2009 JiM9150-Egypt-2009 JIM10174-Egypt-2009 JIM10172-Egypt-2008 JiM9151-Egypt-2009 JIM10171-Egypt-2009 JIM10170-Egypt-2009 JIM10169-Egypt-2009 JIM10173-Egypt-2009 JIM10119 Inde-fromage traditionnel-2004 JIM9139 Yaourt Egypt-2008 JIM10178-Egypt-2009 JiM9154-Egypt-2009 CNRZ1595 Autriche-Emmental-1979 CNRZ778-levain lactique a yaourt-bulgari JIM10155 lait de traite-2009 (C5) JIM1004-Bulgaria-environment CNRZ759 Bulgarie-LevainCY-1978 LMG18311 UK-Yaourt-1974 CNRZ385 Japon-Yaourt sucre-1971 CNRZ406 levain lactique a yaourt CNRZ389 Levain yaourt-1971 CNRZ403-levain lactique a yaourt-1971 CNRZ446 Ivry sur seine-yaourt-1973 CNRZ386-levain lactique a yaourt-1971 JIM10093 Haute Savoie-Lactoserum-1984 CNRZ388 LevainCY-1971 JIM7455 Levain CNRZ1066 LevainCY-1986 CNRZ404-levain lactique a yaourt-1971 CNRZ1588 Italie-fromage CNRZ160 levain lactique commercial a yao CNRZ779 levain lactique Boll 1979 CNRZ1447 Inde Dahli-Lait i fermente-1990 JiM9160-Egypt ,001 A Phylogenetic tree based on concatenated sequences of 6 housekeeping genes in S. thermophilus strains
13 JIM Levain naturel-1999 JIM10076 Haute Savoie-Lactoserum um-1985 JIM10039 Haute Savoie-Levain nat aturel-200 JIM10034 Haute Savoie-Levain natu urel-200 JIM10022 Haute Savoie-Pate Presse see Cuite JIM Haute Savoie-Levain na aturel-200 JIM10042 Haute Savoie-Levain natu urel-200 JIM10033 Haute Savoie-Levain natu urel-200 JIM10026 Haute Savoie-Pate Press see Cuite JIM10037 Haute Savoie-Levain natu urel-200 JIM10050 Haute Savoie-Levain nature rel-200 JIM10054 Haute Savoie-Levain natu urel-200 CNRZ456 levain lactique a gruyere de Com JIM10014 Haute Savoie-Pate Pressee Cuite JIM Haute Sav avoie-pate Pressee Cuit JIM10047 Haute Savoie-Levain naturel-200 JIM10085 Haute Savoie-Lactoserum-1986 CNRZ24B Jura-L -Levain Comte-1959 JIM10104 Levain naturel-1999 JIM10112 Levain naturel-1999 JIM10156 Fro omage (C5) JIM10055 Isere-La Lactoserum-1985 CNRZ1120 Leva vain Comte-1987 CNRZ457-levain artisanal a g gruyere de co JIM Haute Savoie ie-lactoserum-1984 JIM Savoie-Lacto ctoserum-1985 JIM 10064Savoie-Lacto ctoserum-1984 JIM Savoie-Lac Lactoserum-1985 JIM Lait cru-2001 JIM10099 Lait cru-2001 ia-1962 JIM10176-Egypt-2008 JIM10123 pate molle JIM10120 pate molle-2006 CNRZ1579-Itali JIM Haute Savoie-Pate Pressee Cuit JIM10021 Haute Savoie-Pate Pressee Cuite JIM10013 Haute Savoie-Pate Pressee Cuite JIM10012-Bulgaria-environment JIM10121 fromage traditionnel-2004 JiM9156-Egypt-2009 JIM10153 Environnement-2009 (C5) JIM10359 fromagerie-2008 (C3) CNRZ1584-Italia-1962 JIM10151 sol-2009 (C4) CNRZ1587 JIM10175-Egypt-2009 CCHSS9 Hospital-2002 CNRZ1691 Irelande-LevainCY-1992 JIM10254 trayon Toupie (C1) LMD9 U USA-fromage JIM10031 Haute Savoie-Lait CNRZ302-levain artisanal a gruyere de co CNRZ310 Barretaine-levain artisanal a gr JIM10300 Fromage J+2 2nde Campagne (JIM CNRZ701Finlande Helsinki-levain a emment CNRZ703 Mongolie-Lait fermente-1974 CNRZ1592 Allemagne-LevainCY-1987 JIM8232 Lait-2002 JIM10118 ferment-2000 JIM 7443 Levain - JIM 7442 Levain - JIM 7444 Levain (JIM1 JIM10072 Hau aute Savoie-Lactoserum JIM10010 Haute Savoie-Pate Pressee Cuite JIM10097 fromage frai ais-1999 JIM10016 Haute Savoie-Pate Pressee Cuit te JIM10052Haute Savoie-Levain naturel JIM10018 Haute Savoie-Pate Pressee Cuit uite JIM10058 Haute Sao one-lactoserum-1988 JIM10078 Haute Sav avoie-lactoserum-1986 JIM10139 ferment yaourt (C2) JIM Haute Savoi oie-pate Pressee Cuit JIM10087 Haute Savoie-Lactoser erum-1984 JIM10152 lait traite-fromage ge-2009 (C4) JIM10154 lait de traite (C5) JIM10256 Ferment yaourt (C1) JIM lait empresurage (C3) JIM10096 fromage frais-1999 JIM Egypt-2008 CNRZ Italie-frompm-1962 TINA Spa pata 86 TINA Sp pata 64 TINA Spata 66 TINA Labichino 110 TINA Spata 71 CNRZ22-levain artisana nal a gruyere de com TINA Lissandrello 19 TINA Sammito 44 TINA Sammito32 JIM10116 Slovaq TINA Sammito 56 TINA Spata 88 TINA Spata 93 CNRZ779 leva vain lactique Boll 1979 CNRZ Italie-fromage CNRZ160 levain lactiq que commercial a yao JIM7455 Levain 2 1 CNRZ1066 LevainCY-1986 CNRZ404-levai ain lactique a yaourt-1971 CNRZ388 LevainCY CNRZ389 Levain yaourt urt-1971 CNRZ levain lactique a yaourt LMG18311 UK-Yao aourt-1974 JIM10093 Hau ute Savoie-Lactoserum CNRZ Ivry sur seine-yaourt CNRZ4 Z403-levain lactique a yaour urt-1971 CNRZ385 Japon-Yaourt suc ucre-1971 JiM Egypt-2009 CNRZ RZ406 levain lactique a yaour urt CNRZ1595 Autriche-Emmental-1979 JIM1004-Bulgaria-envi nvironment CNRZ759 Bulgarie-LevainC ncy-1978 JIM9139 Yaourt Egypt-2008 JiM9160-Egypt-2 pt-2009 JIM10 M10177-Egypt-2009 JIM10155 lait de traite-2 e-2009 (C5) JIM Egypt-2009 CNRZ778-levain lactique a yaourt-bulgari JIM10173-Egypt-2009 JIM10119 Inde e-fromage traditionnel-2004 CNRZ1447 Inde Dahli-Lait fermen mente-1990 JIM7446 Levain JIM10180-Egypt-2009 JIM10172-Egypt-2008 JIM10174-Egypt-2009 JiM9150-Egypt-2009 JIM10169-Egypt-2009 JiM9151-Egypt-2009 JIM10170-Egypt-2009 JIM10171-Egypt-2009 e frais-1999 JIM10089 Haute Savoie-Lactoserum-1985 JIM10059 Savoie Savoie-Lactoserum Lactoserum JIM10032 Haute Savoie-Lait-1999 JIM10024 Haute Savoie-Pate Pre Pressee Cuite 198 CNR NRZ1202 Grece Heraclion-Yaourt grec-198 CNRZ /JIM8392 Grece Athenes-Yaourt gre CNRZ1 RZ1213 Grece Heraclion-Yaourt grec-198 CNRZ27-bioghurt-1960 JIM10098 fromage frai JIM10106 Levain naturel 19 vain naturel-1999 ge frais-1999 CNRZ1593 CNRZ1237 CNRZ CNRZ368 NL-LevainCY JIM1 M10010-Bulgaria-environm ment-1998 JIM10011-Bulgaria-e environment JIM10013-Bulga aria-environment JIM10124 pate molle-2006 JIM1009-Bulgaria-en nvironment-1998 JIM1003-Bulgari ria-environment JIM1006-Bulgar aria-environment JIM10014-Bulga aria-environment CNRZ1157-Grece Heraclion-Yaourt grec-19 JIM10095 fromage frais CNRZ1156-Crete-Yaourt grec-1987 CNRZ1208-Crete-Yaourt grec-1987 CNRZ308-levain artisa sanal a gruyere de co CNRZ26-levain artisanal agruyere de com JIM1002-Bulgari ria-environment-1997 JIM1001-Bulg lgaria-environment-1997 CNRZ785-lev evain lactique-1979 JIM10117 Inde-fromage trad aditionnel-2004 CNRZ761-levain lactique a yaourt-1978 CNRZ21 Lemuy- y-levain artisanal a gruyer ere JIM Bulgaria-environment CNRZ758 Bulgarie ie-levaincy-1978 CNRZ447-Netherland d-yaourt 1973 JIM Bulgaria-environment JIM1007-Bulg ulgaria-environment CNRZ409 leva vain lactique commercial TINA Sammito 38 quie-fromage traditionnel
14 Industrial Traits of S. S thermophilus hil Strains Test Lactose Galactose Urease Exopolysaccharide fermentation fermentation Production Production No of positive strains (%) 70 (100%) 27 (40%) 5 (10%) 3 (5%) No of Negative strains (%) (0%) (60%) (90%) (95%) Total
15 Conclusions - A discrete level of genetic and phenotypic diversity does exist among S. thermophilus isolated from traditional Egyptian dairy products.
16 Acknowledgments - This work has been carried out in collaboration between the FSMP lab at Mansoura University and the French Laboratoire de Génétique Microbienne lab at INRA within the context of the Egypt-France Science and Technology Cooperation Programme (IMHOTEP).
17 Thanks
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