isolation, molecular characterization and probiotic potential of … · 2019. 7. 30. · isolation,...

Research ArticleIsolation Molecular Characterization and Probiotic Potential ofLactic Acid Bacteria in Saudi Raw and Fermented Milk

Maged S Bin Masalam1 Ahmed Bahieldin1 Mona G Alharbi1 Saad Al-Masaudi1

Soad K Al-Jaouni2 Steve M Harakeh 3 and Rashad R Al-Hindi1

1Department of Biology Faculty of Science King Abdulaziz University Jeddah Saudi Arabia2Department of Pediatric HematologyOncology Yousef Abdullatif Jameel Chair of Prophetic Medicine ApplicationKing Abdulaziz University Hospital Faculty of Medicine KAU Saudi Arabia3Special Infectious Agents Unit King FahdMedical Research Center Yousef Abdullatif Jameel Chair of PropheticMedicine ApplicationKAU Saudi Arabia

Correspondence should be addressed to Steve M Harakeh sharakehgmailcom

Received 28 December 2017 Revised 22 March 2018 Accepted 12 July 2018 Published 25 July 2018

Academic Editor Mohammed S Ali-Shtayeh

Copyright copy 2018 Maged S Bin Masalam et al This is an open access article distributed under the Creative Commons AttributionLicense which permits unrestricted use distribution and reproduction in any medium provided the original work is properlycited

Probiotic bacteria can confer health benefits to the human gastrointestinal tract Lactic acid bacteria (LAB) are candidate probioticbacteria that are widely distributed in nature and can be used in the food industry The objective of this study is to isolate andcharacterize LAB present in raw and fermented milk in Saudi Arabia Ninety-three suspected LAB were isolated from thirteendifferent types of raw and fermented milk from indigenous animals in Saudi Arabia The identification of forty-six selected LABstrains and their genetic relatedness was performed based on 16S rDNA gene sequence comparisons None of the strains exhibitedresistance to clinically relevant antibiotics or had any undesirable hemolytic activity but they differed in their other probioticcharacteristics that is tolerance to acidic pH resistance to bile and antibacterial activity In conclusion the isolates LactobacilluscaseiMSJ1 Lactobacillus caseiDwan5 Lactobacillus plantarum EyLan2 and Enterococcus faeciumGail-BawZir8 are most likely thebest with probiotic potentials We speculate that studying the synergistic effects of bacterial combinations might result in a moreeffective probiotic potential We suspect that raw and fermented milk products from animals in Saudi Arabia especially Labanmade from camel milk are rich in LAB and have promising probiotic potential

1 Introduction

Probiotic bacteria can confer health benefits to the humangastrointestinal tract [1ndash3] Lactic acid bacteria (LAB) arecandidate probiotic bacteria [4] that are widely distributed innature and can be used in the food industry [5] LAB are agroup of microaerophilic or anaerobic Gram-positive bacte-ria that are unable to form spores or produce catalase and arecharacterized by the absence of the cytochrome system [6 7]and the ability to produce antimicrobials for biopreservation[8 9] Certain foods including dairy products for exampleyogurt are considered good sources of probiotics [10] Themajority of microbiota in rawmilk and fermentedmilk prod-ucts include the genera Lactobacillus Enterococcus Lactococ-cus Leuconostoc Pediococcus Oenococcus Carnobacterium

Streptococcus and Weissella [11ndash13] The most obvious ben-efits of LAB fermentation include increased food palatabilityand improved shelf life [14] LAB are generally recognized assafe (GRAS) because they are able to produce bacteriocinsand their consumption confers several health benefits suchas controlling intestinal infections improving lactose uti-lization lowering blood ammonia levels providing efficientresistance against gastric acid and bile [11 15ndash19] influencingthe immune system and lowering serum cholesterol levels[20 21] LAB also adhere to the gastrointestinal tract andconfer pathogen inhibition [11 22 23] Interestingly thepresence of LAB resulted in no change or small changes inthe abundance of other intestinal microbial groups [24]

DNA-based molecular identification of the 16S rRNAgene can discriminate between closely related bacterial

HindawiEvidence-Based Complementary and Alternative MedicineVolume 2018 Article ID 7970463 12 pageshttpsdoiorg10115520187970463

2 Evidence-Based Complementary and Alternative Medicine

species [25ndash29] Amplified ribosomal DNA restriction anal-ysis (ARDRA) and randomly amplified polymorphic DNA(RAPD) were successfully used in discriminating LAB [3031] The approaches include PCR-RFLP followed by directsequencing of the 16S rDNAgene and a BLAST search againstthe sequences of other organisms that are available at theNational Center for Biotechnology Information (NCBI) [32]

The isolation identification and characterization of novelLAB strains have two benefits The first is to reveal thecharacteristic taxonomy of the LAB and the second is toobtain promising beneficial and functional probiotic LAB[33 34] There is little research regarding the isolation andcharacterization of LAB from dairy products Therefore theaim of the present work was to isolate and to identify at themolecular level the lactic acid bacteria contained in rawmilkand fermented milk which were produced indigenously inSaudi Arabia These LAB were evaluated for their functionaltraits probiotic properties and ability to inhibit the growthof pathogenic and food poisoning bacteria

2 Materials and Methods

21 Samples andMedia Thirteen (05 kg) samples of rawmilkand traditional fermented milk from indigenous animalsprocured in Jeddah Province Saudi Arabia were used in thisstudy These samples were collected from the local marketand were stored in a fridge until use MRS agar and MRSbroth media (Oxoid119879119872 Thermo Fisher Scientific USA) wereused to isolate and support the growth of LAB and to inhibitthe growth of unwanted bacteria [35] MRS agar and brothwere also used to enumerate the LAB M17 agar and M17broth (Oxoid Thermo Fisher Scientific USA) were used forisolating the streptococci in dairy products [36 37] Bloodagar (Oxoid Thermo Fisher Scientific USA) and Muller-Hinton agar media (MHA) (HiMedia India) were used toevaluate hemolytic activity and antimicrobial activity of theLAB respectively

22 Isolation of LAB and Maintenance Method 50 g of thesamples was placed into sterile stomacher bags and then werediluted (110) with MRS broth processed to enrich the LABby stomaching sealed and incubated overnight at 30∘C [38]Lactic acid bacteria were isolated by 5-fold serial dilutionsAt the beginning 1mL of sample stomachate was added to9mL of sterile physiological water (085 NaCl) and wasfurther serially diluted Then 01mL aliquots of the samplesrsquosuitable dilutions were plated onto MRS agar and M17 agarTheplateswere incubated at 37∘C for 24-48 h in anaerobic jarsusing the AnaeroGen anaerobic system (Oxoid ThermoFisher Scientific USA) [25 39 40] and in microaerobicconditions (5 CO2) [41] MRS agar plates were used toenumerate the initial growth of the LAB in each sample [25]All experiments were performed in triplicate The resultingisolates were randomly selected from the medium surfaceand were streaked twice on fresh MRS to be purified [35]The isolated strains were maintained by culturing them inMRS broth medium at 30∘C and then storing at minus80∘C inMRS broth containing 50 (vv) glycerol MRS broth andM17 broth were used in all subsequent experiments [42 43]

Isolates from the stocks were subcultured in MRS broth forfurther studies

23 Phenotypic and Genotypic Identifications For all ofthe isolated strains the colony morphology on MRS solidmedium and M17 agar was determined visually and theirmotility was recorded using the hanging drop techniqueGram staining was performed to determine the cell mor-phology and Gram stain reaction of the isolates Cata-lase and cytochrome oxidase tests were also performed[39 42] As for genotypic identification cell pellets wereharvested from 2mL of overnight cultures (up to 2 times109 bacterial cells) of LAB grown in MRS broth by cen-trifugation for 10min at 5000 timesg and the supernatantwas discarded Then DNA extraction was done using aGeneJET Genomic DNA Purification Kit (cat no K0721Thermo Fisher Scientific Waltham MA USA) following themanufacturerrsquos instructions PCR was carried out and theuniversal primers were designed and synthesized (Medicbio-trade Germany) to amplify nearly the entire region of the16S rDNA gene the primers were 16SF27 (forward 51015840-AGAGTTTGATCCTGGCTCAG-31015840) and 16SR27 (reverse51015840-AAGGAGGTGATCCAGCCGCA-31015840) The primers weredesigned to amplify the 16S rDNA gene (sim16 kb [44])with GC contents of 50-60 for a higher melting temper-ature (Tm) to avoid nonspecific amplification [39] PCRconditions were one cycle of 5min of initial denaturationat 95∘C followed by 35 cycles of amplification Each cycleconsisted of denaturation at 95∘C for 30 sec annealing at58∘C for 30 sec and extension at 72∘C for 1min The lastcycle was 10min at 72∘C as a postextension step Elec-trophoresis in 05x TBE buffer was conducted on an 08agarose gel for the amplicons with a 1 kb DNA standard(TrackIt Invitrogen Waltham MA USA) and the gelswere stained with ethidium bromide and photographed Theamplicons were purified they were sequenced for the 16Sgene at Macrogen Inc (Seoul Korea) and they were BLAST-searched to detect similar sequences in the NCBI database(httpswwwncbinlmnihgov) using the Discovery StudioGene v15 (DSGene v15) program [32 42] A fragment ofsim1000 nt involving five variable (V) regions was further uti-lized Binary data metrics were entered into TFPGA (version13) and were analyzed using a qualitative route to generatea similarity coefficient The dissimilarity coefficients wereused to construct dendrograms by the sequential hierarchicaland nested clustering (neighbor joining (NJ)) method withNTSYSpc (version 210 Exeter Software)

24 Antibiotic Susceptibility Hemolytic Activity and AcidBile Tolerance The antibiotic susceptibility of the identi-fied LAB was determined by using the agar disk diffusionmethod [45] Fifteen antibiotics (Oxoid Thermo Fisher Sci-entific USA) were selected as representatives of the differentclasses of clinically important antibiotics Inoculates (106CFU mLminus1) of the LAB were swabbed onto the surfacesof Muller-Hinton agar plates and then the disks of antibi-otics (amoxicillin25120583g penicillin G10 units ceftriaxone30 120583g cefoxitin30120583g tobramycin10120583g oxacillin1120583g bac-itracin10 units chloramphenicol30120583g polymyxin B300

Evidence-Based Complementary and Alternative Medicine 3

units gentamycin10120583g neomycin30120583g clindamycin2120583gvancomycin256120583g nitrofurantoin300120583g and nalidixicacid30 120583g) were applied onto the surface of the platesand incubated at 37∘C and 5 CO2 for 24 h [17 46] Theplates were examined for the presence of inhibition zonesaround the antibiotic disks [43] The inhibitory effect ofthe antibiotics was expressed as the diameter (mm) of theinhibition zones

Bacterial cells were grown on Columbia blood agar(Oxoid Thermo Fisher Scientific USA) supplemented with5 (vv) human blood (obtained from King Abdulaziz Uni-versity Hospital Jeddah KSA) to determine their ability toproduce different types of hemolysins Plates were incubatedat 37∘C in anaerobic jars (Oxoid Thermo Fisher ScientificUSA) with gas-generating kits After 24-hour and 72-hourincubation periods the results were recorded A clear zoneon the blood agar plates was considered a positive result [42]

Tolerance of the LAB to acid and bile was determined asdescribed [47] All strains were grown at 37∘C for 24 h inMRSbroth and were suspended to an approximate cell concentra-tion of 108 CFU mLminus1 (using turbidity standard McFarland05) in MRS broth adjusted to pH 30 for 2 h and the cellswere then incubated in 05wv bile (Sigma-Aldrich France)for 4 h These conditions were chosen to represent the timerequired for the bacteria to pass through the gastrointestinalsystem and the pH value and bile concentration found inthe stomach and intestine respectively Bacterial viability wasassessed by enumeration onMRS agar plates at zero time andat the end of incubation

25 Evaluation of Antibacterial Activity of Identified StrainsThe agar well diffusion method was used to adequatelyinvestigate the antibacterial potential of the 46 identified LABas described by Schillinger et al (1996) Chahad et al (2012)and Messaoudi et al (2012a) Suspensions of seven indica-tor bacterial cultures namely Staphylococcus aureus ATCC25923 methicillin-resistant Staphylococcus aureus (MRSA)ATCC 43330 Escherichia coli ATCC 25922 Salmonella sppShigella sonnei ATCC 25931 Enterococcus faecalis ATCC29212 and Listeria monocytogenes ATCC 13932 providedby the Microbiology Laboratory of King Abdulaziz Univer-sity Hospital Jeddah KSA were prepared using turbiditystandard McFarland 05 Then 150 120583L of these suspensionswas inoculated onto MHA medium by the streaking platemethod and four wells were punched in each inoculated agarmedium plate using a sterile cork borer (6mm diameter)Cell-free supernatant (CFS) of LAB grown in MRS wasprepared by centrifugation at 10000 timesg for 20min and4∘C Then 100120583L aliquots of the CFS of each strain werepipetted into their designated wells on the plates Finallythe susceptibility of the test organisms against MRS brothwas taken as a control All inoculated plates were incubatedat 37∘C for 18-24 h After incubation the diameter of eachformed inhibition zonewasmeasured twice using ruler thenthe average was taken to represent the antibacterial activity

26 Statistical Analysis Data were expressed as the meanplusmn standard deviation (SD) calculated over independentexperiments performed in triplicate For inference statistics

Figure 1 PCR amplification of the 16S rDNA from the isolated LABon 08 agarose gel M 1 kb DNA marker (TrackIt Invitrogen)Numbers 2-4 7-12 19 23 28 29 32 34-40 42 44-47 50 53 58 6061 63 68 73-83 92 and 93 refer to the strains numbers in Table 1

one-way analysis of variance (ANOVA) was applied and itwas performed with the Statistical Package for the SocialSciences (SPSSPC version 200)

3 Results

31 Isolation and Characterization of LAB A total of 93lactic acid bacteria (LAB) were isolated from the 13 collectedsamples of locally produced raw milk and fermented milksuch as fresh raw milk frozen raw milk fresh cheesesalty cheese cooked cheese stirred yogurt (Laban) qeshta(cream)madheer (dried fermentedmilk) yogurt and butterThese samples originated from different animal sourcesincluding cows goats and camels The LAB counts undermicroaerobic incubation conditions were higher than thoseunder anaerobic conditions For example the mean counts(per gram of sample) in the samples of qeshta made ofcow milk and stirred yogurt ldquoLabanrdquo made of camel milkgrown for 48 h under microaerobic incubation conditionswere between 41 times 106 plusmn 02 times 106 and 22 times 1010 plusmn 02 times1010 CFU gminus1 respectively while these counts were between47 times 102 plusmn 06 times 102 and 27 times 1010 plusmn 03 times 109 CFU gminus1respectively under anaerobic incubation conditions

The colony morphologies of the isolates were visuallyobserved on the surface of MRS solid medium the colorvaried from white to pale creamy the shape was circular andthe size ranged from 05 to 4mm in diameter Most strains(9247) were Gram-positive The isolated strains differedin their catalase and cytochrome oxidase activity where 71were negative for catalase activity and 72 were negative forcytochrome oxidase production activity The motility of theisolated strains also differed when tested by the hanging droptechnique with 96 of the isolates being nonmotile

The 16S rDNAgene (sim16 kb) was amplified from all of theisolated strains (Figure 1)Then the amplicons were column-purified and sequenced The resultant nucleotide sequenceswere BLAST-searched for homology with known sequencesin the NCBI database The results in Table 1 indicated thatthe nucleotide sequences of 46 strains aligned with the16S rDNA sequences of 14 different species belonging tofive genera namely Enterococcus Lactococcus LactobacillusStreptococcus andWeissella (Figure 2) These strains showedidentities of 99 (12 strains) 98 (14 strains) 97 (12strains) 96 (6 strains) and 95 (2 strains) Analysis of the


Table1Genotypes

ofthes

elected46

isolated

LABas

16SrD

NAgene

sequ

encesa

lignm

entssubm

itted

totheN

CBIG

eneB

ankdatabase

Strain

no

Strain

name

GeneB

ankAc

cNo

Query

coverage

Maxident

Isolates

ourcea

2En

terococcus

faecium

Rashad3

KU324893

94

99

RCM

3En

terococcus

durans

Etim

ad1

KU324894

65

98

RCM

4En

terococcus

faecium

SMBM

3KU

324895

98

99

RCM

7La

ctobacillus

caseiM

SJ1

KU324896

98

99

CMCM

8La

ctobacillus

paracaseiN

MBM

1KU

324897

98

99

CMCM

9La

ctobacillus

caseiB

gShn

3KU

324898

99

99

CMCM

10La

ctobacillus

caseiM

asaLam

7KU

324899

95

99

CMCM

11La

ctobacillus

caseiD

wan5

KU324900

96

97

CMCM

12La

ctobacillus

plantarum

EyLan2

KU324901

89

97

CMCM

19La

ctobacillus

futsa

iiEM

BM2

KU324902

99

98

CCMCM

23Weis

sella

confusaSaEd

-7KU

324903

95

98

CCMCM

28En

terococcus

faecalisShbam40

KU324904

99

97

RGM

29En

terococcus

faecium

ZiNb3

KU324905

95

97

RGM

32En

terococcus

faecium

Gail-B

awZir8

KU324906

93

97

SYMCM

34En

terococcus

faecium

NSJ2

KU324907

99

96

SYMCM

35Streptococcustherm

ophilusB

inSlman8

KU324908

99

99

SYMCM

36La

ctococcuslactis

HadRa

mi9

KU324909

98

98

SYMCM

37En

terococcus

faecium

Marwh2

KU324910

97

99

QMCM

38En

terococcus

faecium

Mon

a3KU

324911

99

99

QMCM

39En

terococcus

faecium

SSJ3

KU324912

98

97

QMCM

40En

terococcus

lactisJedd

ah9

KU324913

97

98

QMCM


ophilusM

aNaL33

KU324914

97

98

QMCM

44En

terococcus

faeciumAd

eb3

KU324915

97

98

MMGM

45En

terococcus

faecium

ESJ4

KU324916

99

96

MMGM

46En

terococcus

durans

She7R

KU324917

96

97

MMGM

47La

ctobacillus

acidophilusH

adhram

aut4

KU324918

91

98

MMGM

50La

ctobacillus

acidophilusM

usallam2

KU324919

99

97

YMCM

53En

terococcus

faecium

BagH

om4

KU324920

99

99

RCAM

58Streptococcusequinus

Omer9

KU324921

95

97

RCAM


Anw

r4KU

324922

77

96

SYMCA

M61

Streptococcusequinus

Zaki1

KU324923

98

98

SYMCA

M63


Salam7

KU324924

87

96

SYMCA

M68

Enterococcus

faecium

Ma7Fo

dKU

324925

98

98

SCMCM


JmaL3

KU324926

96

96

FCAM

74S treptococcusequinus

Foad7

KU324927

95

96

FCAM

75Weis

sella

confusaAhM

d8KU

324928

99

99

BMCM

76Weis

sella

confusaTarim

4KU

324929

99

97

BMCM

77Weis

sella

confusaNoo

R1KU

324930

78

95

BMCM

78Weis

sella

confusaSaYu

n2KU

324931

98

98

BMCM

79Weis

sella

confusaMuK

alla5

KU324932

94

95

BMCM


Table1Con

tinued

Strain

no

Strain

name

GeneB

ankAc

cNo

Query

coverage

Maxident

Isolates

ourcea

80Weis

sella

confusaSYary1

KU324933

98

97

BMCM

81Weis

sella

confusaSho7ir

KU324934

96

98

BMCM

82Weis

sella

confusaFarag8

KU324935

97

97

BMCM

83Weis

sella

confusaA7G

afKU

324936

99

98

BMCM

92La

ctococcusgarvieaeZ

SJ5

KU324937

99

98

BMCM

93La

ctococcusgarvieaeE

mad4

KU324938

98

99

BMCM

a RCM

raw

cowmilk

RGMraw

goatmilk

RCA

Mraw

camelmilk

FCA

Mfrozencamelmilk

CMCM

cheesem

adefrom

cowmilk

CCM

CMcoo

kedcheese

madefrom

cowmilk

SCM

CMsalty

cheese

made

from

cowmilk

SYM

CMstirredyogu

rt(Laban)m

adefrom

cowmilk

SYM

CAMstirredyogu

rt(Laban)m

adefrom

camelmilk

YMCM

yogurtm

adefrom

cowmilk

BMCM

butterm

adefrom

cowmilk

QMCM

qeshta(cream

)madefrom

cowmilk

MMGMm

adheer

madefrom

goatmilk


Figure 2 Phylogenetic neighbor joining (NJ) tree based on the 16S rDNA full-length sequences (sim16 kb) of the 46 selected LAB strains ofrawmilk and fermented milk samples based on the results of sequence alignmentThe numbers on the tree refer to the LAB strains in Table 1

16 strains aligned with the genus Enterococcus indicated that12 strains (ie Rashad3 SMBM3 ZiNb3Gail-BawZir8NSJ2Marwh2Mona3 SSJ3 Adeb3 ESJ4 BagHom4 andMa7Fod)might belong to the species E faecium while two strains(ie Etimad1 and She7R) one strain (ie Jeddah9) and onestrain (ie Shbam40) might belong to the species E duransL lactis and E faecalis respectively In studying the threestrains of the genus Lactococcus the results indicated that onestrain (ie HadRami9) might belong to the species L lactiswhile two strains (ie Emad4 and ZSJ5) might belong to thespecies L garvieae Analysis of the nine strains of the genusLactobacillus indicated that two strains (ie Hadhramaut4and Musallam2) four strains (ie MSJ1 BgShn3 MasaLam7and Dwan5) one strain (ie NMBM1) one strain (ieEyLan2) and one strain (ie EMBM2) might belong to thespecies L acidophilus L casei L paracasei L plantarumand L futsaii respectively Two out of the eight strains ofthe genus Streptococcus (ie BinSlman8 andMaNaL33)mightbelong to the species S thermophilus while the rest (ieOmer9 Anwr4 Zaki1 Salam7 JmaL3 and Foad7) mightbelong to the species S equinus In addition all ten strainsof the genus Weissella (ie SaEd-7 AhMd8 Tarim4 NooR1SaYun2 MuKalla5 SYary1 Sho7ir Farag8 and A7Gaf) mightbelong to the species W confusa

32 Susceptibility of LAB to Antibiotics andHemolytic ActivityThe tested 46 LAB underwent an antibiotic susceptibilitytesting and their growthwas inhibited to some extent by themajority of the 15 tested antibiotics (Table 2) Interestingly

three antibiotics (bacitracin gentamicin and neomycin)had a great (sim100) inhibition effect against all the testedstrains with a spectrum of inhibition zones of 140 plusmn 00- 285 plusmn 071 95 plusmn 071 - 400 plusmn 00 and 95 plusmn 071 -315 plusmn 071mm respectively On the other hand penicillinG tobramycin and vancomycin inhibited the growth witha spectrum of inhibition zones of 150 plusmn 141 - 400 plusmn 141110 plusmn 141 - 235 plusmn 071 and 95 plusmn 071 - 235 plusmn 071mm atlower levels of 89 739 and 674 inhibition effect on thetested strains respectively There were only four antibiotics(ie nalidixic acid polymyxin B oxacillin and cefoxitin)that had low inhibition action at levels of 174 304348 and 457 respectively The actions of the antibioticsoxacillin cefoxitin and nalidixic acid were resisted by manystrains of lactobacilli and W confusa Five strains (namelyS thermophilus BinSlman8 S thermophilus MaNaL33 Sequinus Omer9 S equinus JmaL3 and S equinus Foad7)were resistant to penicillin G while 15 strains (326) wereresistant to vancomycin These strains belong to L casei (4strains) L paracaseiNMBM1 L plantarum EyLan2 L futsaiiEMBM2 andW confusa (8 strains)

In our study 32 (6957) of the tested strains were non-hemolytic (120574-hemolysis) while the remaining 14 (3043)strains exhibited 120572-hemolytic activity and these were S ther-mophilus BinSlman8 S thermophilus MaNaL33 W confusaSaEd-7 W confusa AhMd8 W confusa Tarim4 W confusaNooR1W confusa SaYun2W confusaMuKalla5W confusaSho7ir W confusa Farag8 W confusa A7Gaf W confusaSYary1 L garvieae ZSJ5 and L garvieae Emad4 (Table 2)


Table2Th

eprofi

leof

theselected

LAB(n

=46

)for

mainprob

iotic

characteris

ticsa

ndtheantib

acteria

lactivity

spectraof

theirC

FSagainstseven

indicatoro

rganism

sAE

nterococcus

faecalisAT

CC29212B

EcoliAT

CC25922C

Salm

onellasppD

Shigella

sonn

eiAT

CC25931E

Staphylococcus

aureus

ATCC

25923F

MRS

AAT

CC43330

GL

isteria

monocytogenes

ATCC

13932minusnoinhibitio

n+

inhibitio

nzone

80ndash

100mm+

+inhibitio

nzone

101ndash

150mm+

++inh

ibition

zone

151ndash

200mm+

+++

inhibitio

nzone

200ndash300m

m

Strain

no-

code

Samples

ource

Strain

name

Accessionno

Hem

olysis

viabilityin

pH3medium

a

viabilityin

05

wv

bilea

Noof

antib

iotic

ssensitive

to15

AB

CD

EF

G

2RC

M2

Enterococcus

faecium

Rashad3

KU324893120574-hem

olysis

981plusmn0141

988plusmn0141

12+

++minus

+minus

++

3RC

M3

Enterococcus

durans

Etim

ad1

KU324894120574-hem

olysis

994plusmn0141

9775plusmn0354

12+

++minus

+minus

++

4RC

M4

Enterococcus

faecium

SMBM

3KU

324895120574-hem

olysis

991plusmn0283

981plusmn0283

13+

++minus

+minus

++

7CM

CM1

Lacto

bacilluscaseiMSJ1

KU324896120574-hem

olysis

9945plusmn0354

9905plusmn0212

9+

++++

+++

++

8CM

CM2

Lactobacillus

paracaseiN

MBM

1KU

324897120574-hem

olysis

9355plusmn0636

9845plusmn0212

10+

+++

+minus

+++

9CM

CM3

Lacto

bacilluscaseiBg

Shn3

KU324898120574-hem

olysis

9885plusmn0919

9925plusmn0212

9+

++

+minus

++

10CM

CM4

Lacto

bacilluscaseiMasaLam

7KU

324899120574-hem

olysis

975plusmn0283

974plusmn0424

9+

+++

+minus

+++

11CM

CM5

Lacto

bacilluscaseiDwan5

KU324900120574-hem

olysis

987plusmn0849

993plusmn0283

9+

+++

+++

++

12CM

CM6

Lactobacillus

plantarum

EyLan2

KU324901120574-hem

olysis

763plusmn0566

990plusmn0141

11+

++

++

++

19CC

MCM

1La

ctobacillus

futsa

iiEM

BM2

KU324902120574-hem

olysis

9835plusmn0778

918plusmn12

77

+++

+++minus

+++

23CC

MCM

5Weis

sella

confusaSaEd

-7KU

324903120572-hem

olysis

991plusmn0141

9545plusmn0354

10minus

minus+minusminus

+++

28RG

M5

Enterococcus

faecalisShbam40

KU324904120574-hem

olysis

822plusmn099

6745plusmn16

2612

minus++

++++

minusminus

+++

29RG

M6

Enterococcus

faecium

ZiNb3

KU324905120574-hem

olysis

994plusmn0141

996plusmn0141

12+

+minus

++

+++

32SY

MCM

3En

terococcus

faecium

Gail-B

awZir8

KU324906120574-hem

olysis

991plusmn0283

741plusmn2263

12++

+++

+++

++

34SY

MCM

5En

terococcus

faecium

NSJ2

KU324907120574-hem

olysis

9896plusmn0078

976plusmn0424

11+

++

++minus

+++

++35

SYMCM

6Streptococcustherm

ophilusB

inSlman8

KU324908120572-hem

olysis

6735plusmn14

859565plusmn0636

11minus

++minus

++++

minus+

36SY

MCM

7La

ctococcuslactis

HadRa

mi9

KU324909120574-hem

olysis

9895plusmn0212

655plusmn19

812

minus+

+++

minusminus

++37

QMCM

1En

terococcus

faecium

Marwh2

KU324910120574-hem

olysis

985plusmn0424

8365plusmn12

0212

++minus

+minus

++

38QMCM

2En

terococcus

faecium

Mon

a3KU

324911120574-hem

olysis

9755plusmn0354

6815plusmn16

2611

+++

++minus

+++

39QMCM

3En

terococcus

faecium

SSJ3

KU324912120574-hem

olysis

983plusmn0283

609plusmn12

7312

+++minus

+minus

++

40QMCM

4En

terococcus

lactisJedd

ah9

KU324913120574-hem

olysis

9715plusmn0354

6715plusmn0636

11minus

++

+minus

++

42QMC M

6Streptococcustherm

ophilusM

aNaL33

KU324914120572-hem

olysis

6085plusmn14

85951plusmn0424

11minus

++minus

++++

minus+

44MMGM1

Enterococcus

faecium

Adeb3

KU324915120574-hem

olysis

985plusmn0566

9735plusmn0636

8+

+++

++minus

minus++

45MMGM2

Enterococcus

faecium

ESJ4

KU324916120574-hem

olysis

991plusmn0283

598plusmn0283

12++

+minus

+minus

+++

46MMGM3

Enterococcus

durans

She7R

KU324917120574-hem

olysis

956plusmn0424

667plusmn12

7311

+++minus

+minus

+++

47MMGM4

Lactobacillus

acidophilusH

adhram

aut4

KU324918120574-hem

olysis

8265plusmn0636

907plusmn1131

10minus

+++

++minus

+++

50YM

CM3

Lactobacillus

acidophilusM

usallam2

KU324919120574-hem

olysis

9225plusmn0495

9945plusmn0212

11minus

+++

++minus

+++

+53

RCAM1

Enterococcus

faecium

BagH

om4

KU324920120574-hem

olysis

995plusmn01 41

9825plusmn0495

13+

++minus

+minus

++

58RC

AM6


Omer9

KU324921120574-hem

olysis

6455plusmn2758

975plusmn0283

13minus

++

++minus

minus++

60SY

MCA

M2


Anw

r4KU

324922120574-hem

olysis

7285plusmn12

029745plusmn0495

14minus

++

++minus

+++

++61

SYMCA

M3


Zaki1

KU324923120574-hem

olysis

7575plusmn19

09971plusmn0849

14minus

++

++minus

minus+

63SY

MCA

M5


Salam7

KU324924120574-hem

olysis

7445plusmn0636

9895plusmn0212

14minus

++

++minus

+++

+68

SCMCM

4En

terococcus

faecium

Ma7Fo

dKU

324925120574-hem

olysis

9845plusmn0636

377plusmn0990

13+

+++

++minus

++

73FC

AM4


JmaL3

KU324926120574-hem

olysis

5285plusmn0636

9555plusmn0636

13minus

+minusminus

++

++74

FCAM5


Foad7

KU324927120574-hem

o lysis

5675plusmn14

85953plusmn0566

12minus

+minusminus

++

++75

BMCM

1Weis

sella

confusaAhM

d8KU

324928120572-hem

olysis

9945plusmn0212

953plusmn0424

10minus

minus+minusminus

minusminus

76BM

CM2

Weis

sella

confusaTarim

4KU

324929120572-hem

olysis

9915plusmn0212

952plusmn0424

10minus

minus+minusminus

++++minus

77BM

CM3

Weis

sella

confusaNoo

R1KU

324930120572-hem

olysis

9955plusmn0071

9655plusmn0778

10++

++minusminus

+++minus

78BM

CM4

Weis

sella

confusaSaYu

n2KU

324931120572-hem

olysis

9945plusmn071

9635plusmn0495

10minus

++++minusminus

minusminus

79BM

CM5

Weis

sella

confusaMuK

alla5

KU324932120572-hem

olysis

709plusmn0707

9595plusmn0212

11minus

+minusminus

+++

+


Table2Con

tinued

Strain

no-

code

Samples

ource

Strain

name

Accessionno

Hem

olysis

viabilityin

pH3medium

a

viabilityin

05

wv

bilea

Noof

antib

iotic

ssensitive

to15

AB

CD

EF

G

80BM

CM6

Weis

sella

confusaSYary1

KU324933

120572-hem

olysis

992plusmn0141

982plusmn0424

11minus

+minusminusminusminus

+81

BMCM

7Weis

sella

confusaSho7ir

KU324934

120572-hem

olysis

969plusmn0566

949plusmn0849

10++

minus+

++

++

82BM

CM8

Weis

sella

confusaFarag8

KU324935

120572-hem

olysis

973plusmn0424

9565plusmn0636

10++

minus+minusminusminusminus

83BM

CM9

Weis

sella

confusaA7G

afKU

324936

120572-hem

olysis

9675plusmn0212

9515plusmn0636

10++

minus+minusminus

+minus

92BM

CM18

LactococcusgarvieaeZ

SJ5

KU324937

120572-hem

olysis

885plusmn0707

9865plusmn0354

11minus


93BM

CM19

LactococcusgarvieaeE

mad4

KU324938

120572-hem

olysis

860plusmn0707

992plusmn0141

11minus


a Resultsarep

resented

asthem

eanvalueo

fdup

licatetria

lsplusmnstandard

deviation(SD)


3770 50-70 701-899 90-95 951-979 98-9955

Percentage of viability

No of acidity resistant strainsNo of bile resistant strains

0

5

10

15

20

25

Num

ber o

f str

ains

Figure 3The percentage viability of the 46 LAB grouped accordingto their acidity and bile tolerance

33 Acid and Bile Tolerance The results showed that the46 tested strains tolerated the acidic condition by variableratios The percentage viability of 50-70 701-899 90-95 951-979 and 98-9955 occurred among 5 9 27 and 23 strains respectively (Table 2 and Figure 3) Themost acid-tolerant strains were enterococci such as E fae-cium SMBM3BagHom4 ZiNb3 Gail-BawZir8 ESJ4 NSJ2Marwh2 SSJ3 Etimad1 and other LAB genera such as Wconfusa SaEd-7 AhMd8 Tarim4 NooR1 SaYun2 SYary1 Lcasei MSJ1 BgShn3 Dwan5 L futsaii EMBM2 and L lactisHadRami9 (Table 2) The 46 LAB with acidity tolerancesranging from 5285 to 9955 were further tested for 4 hfor 05 wv bile tolerance The obtained results showed thatthe survival percentages were 377 50-70 701-899 90-95 951-979 and 98-9955 for 1 6 2 3 20 and 14strains respectively (Table 2 and Figure 3) A total of 717of the strains were bile-tolerant without any significant lossof viability (gt95 survival) The most bile-tolerant strainswere among enterococci such asE faeciumZiNb3E faeciumRashad3 and E faecium SMBM3 and other species such asL casei BgShn3 L casei Dwan5 L casei MSJ1 L plantarumEyLan2 L acidophilus Musallam2 L paracasei NMBM1 Sequinus Salam7 L garvieae Emad4 L garvieae ZSJ5 and Wconfusa SYary1 (Table 2)

34 Antibacterial Activity of Isolated LAB Spectra of theantibacterial activity of the CFS preparations for the 46 LABagainst seven bacterial indicator strains (4 Gram-positiveand 3 Gram-negative) are shown in Table 2 There werevariable spectra of inhibition zones of the antibacterialactivity of the identified strains against the indicator bacteriawhich ranged from 8 to 30mm in diameter The resultsindicated antibacterial activity of the CFS of 25 (5435) 38(8261) 33 (7172) 31 (6739) 11 (2391) 33 (7172)and 38 (8261) LAB strains against E faecalis E coliSalmonella spp Shigella sonnei S aureusMRSA and Listeriamonocytogenes respectively L casei MSJ1 L casei Dwan5L plantarum EyLan2 and E faecium Gail-BawZir8 strainsshowed antibacterial activity against all indicator bacteriaat the different recorded spectra of inhibition 20-30mmzones of inhibition activity were noticed for S thermophilus

BinSlman8 and S thermophilus MaNaL33 against S aureusand for W confusa Tarim4 against MRSA 151-20mm zonesof inhibition were recorded for E faecium NSJ2 S equinusAnwr4 S equinus Salam7 and W confusa NooR1 againstMRSA and for E faecalis Shbam40 and L acidophilusMusal-lam2 against Listeria monocytogenes

4 Discussion

Identified strains (68) were Gram-positive catalase-negative and oxidase- and hemolysis-producing as wellas chain-forming fermentative cocci and rods Theseconclusions meet those reached earlier [9 48] in the study onmilk-grown LAB Our pilot study aimed at the identificationof LAB isolated from different types of dairy-based foodsobtained from various animals in order to have a completepicture of the LAB found in those products The resultsconfirmed that the distribution of the isolated LAB issample-dependent Enterococci were recovered from qeshtamadheer raw cow milk and raw goat milk Lactobacilliwere isolated from cheese and yogurt made from cowmilk Streptococci were abundant in stirred yogurt (Laban)made from camel milk and in frozen camel milk andWeissella were isolated from butter made from cow milkA phylogenetic tree was successfully generated from themultiple sequence alignment of full- and partial-length 16SrDNA sequences of the 46 strains We speculate that the LABof the different genera can be positively discriminated usingonly the hypervariable region V2 Our data was alignedwith that generated by Balcazar et al [49] who indicatedthat the sequence containing both the V1 and V2 regionscompletely discriminated among LAB strains Chakravortyet al [50] also indicated that the V1 V3 V5 V6 and V7regions are conserved while the V2 V4 V8 and V9 regionsare hypervariable in the family

The antibiotic resistance of probiotic LAB is a con-troversial subject as they may be reservoirs of antibioticgenes The safety of LAB regarding food applications wasevaluated by screening for the presence of virulence factorscoding genetic determinants and by testing their phenotypicresistance to different antibiotics [51] It is believed thatthe bacteria present in the intestinal microflora of food-producing animals may acquire antibiotic resistance Thenby the exchange of genetic material the antibiotic-resistantbacteria can transfer the resistance factor to other pathogenicbacteria [33 52] To resolve this problem in probiotic studiesit is necessary to certify that a prospective probiotic straincontains no transferable resistance genesThe 15 tested antibi-otics were selected for their variety of action mechanismsrevealing different profiles of resistance for the strains similarto the results of Schirru et al [43] The different enterococcalstrains were sensitive to vancomycin while Chahad et al[42] and Haghshenas et al [46] reported that all testedenterococci were resistant to vancomycin they claimed thatthis resistance is an intrinsic property for most LAB Theisolated L plantarum EyLan2 was sensitive to penicillin Gchloramphenicol gentamicin and clindamycin in agreementwith the results of Haghshenas et al [46] However thepresence of antibiotic resistance properties among probiotic


bacteria is advantageous as this allows the bacteria to survivein the gastrointestinal tract during antibiotic treatment Inour study lactobacilli and all other strains were sensitiveto aminoglycosides represented by gentamicin while veryfew were resistant to clindamycin Among streptococci 100were sensitive to gentamicin and chloramphenicol whileFederici et al [11] reported that 1667 of LAB isolates wereresistant to clindamycin and 6923 and 1538of lactobacilliwere resistant to gentamicin and clindamycin respectivelyand 75 and 75 of streptococci exhibited resistance togentamicin and chloramphenicol respectively

The absence of cytolysin coding genes is a good char-acteristic in the food applications of enterococci and otherLAB Cytolysin is a bacterial toxin expressed by some isolatesof E faecalis which displays both hemolytic and bactericidalactivities [51] All the 46 studied strains exhibited no 120573-hemolytic activity which is in agreement with Chahad etal [42] and Bozoudi et al [8] This characteristic confirmsthat these LAB can be used safely in food applications Theability of the isolated LAB strains to resist acid and bile is animportant probiotic property since they have to survive theconditions in the stomach and the small intestine [53 54]The 46 strains identified in the present study differentiallytolerate the acidic and bile conditions 59 of the isolatedLAB were tolerant at a high rate Washington et al [3] alsoindicated that 72 of the tested LAB are tolerant to acidityand bile at a survival rate of 90 while Messaoudi et al[53] reported that the L salivarius SMXD51 can tolerategastrointestinal conditions (pH 3 acidity and 05 wv bile)with a 99 rate of survival In our study the behavior ofmost lactobacilli especially the strains L caseiMSJ1 L caseiBgShn3 and L Casei Dwan5 was similar to that observedby Messaoudi et al [53] Ladda et al [55] indicated that Lparacasei L casei andW confusawere tolerant to acidity andbile at pH 30 and 4 bile respectively Bujnakova et al [33]indicated that L salivarius L agilis L reuteri L murinusand L amylovorus were tolerant to acidity and bile at pH25 and 03 bile respectively The strains of L paracasei Lcasei and Lactobacillus rhamnosus isolated by Reale et al [2]exhibited high acidity and bile tolerance at pH 35 and 15bile respectively However Ahmadova et al [51] reportedthat E faecium AQ71 could not grow at pH 3 and 4 while itcan grow in the presence of bile concentrations ranging from02 to 03

As for the antibacterial activity the largest antimicro-bial spectrum in the present study was exerted by CFSof Lactobacillus casei MSJ1 Lactobacillus casei MasaLam7Lactobacillus casei Dwan5 Enterococcus faecium BawZir8Lactobacillus paracasei NMBM1 Lactobacillus casei BgShn3and Lactobacillus plantarum EyLan2 as they inhibited allthe indicator strains The indicator strain E coli ATCC25922 in the present study was sensitive to as many as 38LAB strains The LAB isolates studied by Bozoudi et al [8]almost inhibited the growth of E faecalis E coli (100) Saureus and L monocytogenes In our study no inhibitionwas observed by the isolated E faecalis Shbam40 against Efaecalis (ATCC 29212)

Our results indicated that the tested Gram-positiveindicator bacteria were more sensitive to the antimicrobial

activity of LAB to some extent than the Gram-negative oneswhich agrees partially with the results of Ghanbari et al [56]Washington et al [3] indicated no clear relationship betweenthe Gram type of indicator bacteria and their sensitivityto LAB They also stated that the number of isolated LABinhibiting the growth of L monocytogenes (ATCC 7644) wasgreater than the number of those inhibiting E faecalis (ATCC19433) E coli (ATCC 8739) Salmonella Typhi (ATCC 6539)Shigella flexneri (ATCC 12022) and S aureus (ATCC 25923)On the other hand Strompfova and Laukova [24] showedthat the growth of all tested Gram-negative indicators washighly inhibited by LAB compared to the inhibition of Gram-positive indicators

5 Conclusions

The raw and fermented milk of animals from Saudi Arabiaespecially stirred yogurt (Laban) made from camel milk wasconfirmed to be rich in LABThemost important strains withpromising probiotic potential for beneficial applications areLactobacillus casei MSJ1 Lactobacillus casei Dwan5 Lacto-bacillus plantarum EyLan2 and Enterococcus faecium Gail-BawZir8 We argue that studying the synergistic effects ofbacterial combinations might result in the occurrence of amore effective probiotic potential

Conflicts of Interest

The authors declare that there are no conflicts of interest

Acknowledgments

The authors would like to acknowledge Faculty of ScienceKing Abdulaziz University Jeddah Saudi Arabia for theirtechnical and financial support

References

[1] FAOWHO Joint FAOWHOWorkingGroupReport onDraftingGuidelines for the Evaluation of Probiotics in Food LondonFAOWHO Ontario Canada 2002

[2] A Reale T Di Renzo F Rossi et al ldquoTolerance of Lactobacilluscasei Lactobacillus paracasei and Lactobacillus rhamnosusstrains to stress factors encountered in food processing and inthe gastro-intestinal tractrdquo LWT- Food Science and Technologyvol 60 no 2 pp 721ndash728 2015

[3] W L G de Almeida Junior I D S Ferrari J V de Souza C DA da Silva M M da Costa and F S Dias ldquoCharacterizationand evaluation of lactic acid bacteria isolated from goat milkrdquoFood Control vol 53 pp 96ndash103 2015

[4] S D Todorov and B D G de Melo Franco ldquoLactobacillusplantarum Characterization of the species and application infood productionrdquo Food Reviews International vol 26 no 3 pp205ndash229 2010

[5] L Tserovska S Stefanova and T Yordanova ldquoIdentification oflactic acid bacteria isolated from Katyk goats milk and cheeserdquoJournal of Culture Collections vol 3 pp 48ndash52 2002

[6] L M Cintas M P Casaus C Herranz I F Nes and P EHernandez ldquoReview Bacteriocins of Lactic Acid Bacteriardquo


Food Science and Technology International vol 7 no 4 pp 281ndash305 2001

[7] X Lu L Yi J Dang Y Dang and B Liu ldquoPurification of novelbacteriocin produced by Lactobacillus coryniformisMXJ 32 forinhibiting bacterial foodborne pathogens including antibiotic-resistant microorganismsrdquo Food Control vol 46 pp 264ndash2712014

[8] D Bozoudi C Kotzamanidis M Hatzikamari N TzanetakisG Menexes and E Litopoulou-Tzanetaki ldquoA comparison foracid production proteolysis autolysis and inhibitory propertiesof lactic acid bacteria from fresh and mature Feta PDO Greekcheese made at three different mountainous areasrdquo Interna-tional Journal of Food Microbiology vol 200 pp 87ndash96 2015

[9] G Tabanelli C Montanari E Bargossi et al ldquoControl oftyramine and histamine accumulation by lactic acid bacteriausing bacteriocin forming lactococcirdquo International Journal ofFood Microbiology vol 190 pp 14ndash23 2014

[10] M Palomo A M Gutierrez M C Perez-Conde C Camaraand Y Madrid ldquoSe metallomics during lactic fermentation ofSe-enriched yogurtrdquo FoodChemistry vol 164 pp 371ndash379 2014

[11] S Federici F Ciarrocchi R Campana E Ciandrini G Blasiand W Baffone ldquoIdentification and functional traits of lacticacid bacteria isolated from Ciauscolo salami produced inCentral ItalyrdquoMeat Science vol 98 no 4 pp 575ndash584 2014

[12] K Henrıquez-Aedo D Duran A Garcia M B Hengst andM Aranda ldquoIdentification of biogenic amines-producing lacticacid bacteria isolated from spontaneous malolactic fermenta-tion of chilean red winesrdquo LWT- Food Science and Technologyvol 68 pp 183ndash189 2016

[13] L A Nero M R De Mattos M De Aguiar Ferreira Barros MB T Ortolani V Beloti and B D G De Melo Franco ldquoListeriamonocytogenes and Salmonella spp in raw milk produced inBrazil Occurrence and interference of indigenous microbiotain their isolation and developmentrdquoZoonoses and PublicHealthvol 55 no 6 pp 299ndash305 2008

[14] L OrsquoSullivan R P Ross and C Hill ldquoPotential of bacteriocin-producing lactic acid bacteria for improvements in food safetyand qualityrdquo Biochimie vol 84 no 5-6 pp 593ndash604 2002

[15] K Angmo A Kumari Savitri and T C Bhalla ldquoProbioticcharacterization of lactic acid bacteria isolated from fermentedfoods and beverage of Ladakhrdquo LWT- Food Science and Technol-ogy vol 66 pp 428ndash435 2016

[16] T K A Bezerra N M D O Arcanjo A R R D Araujoet al ldquoVolatile profile in goat coalho cheese supplementedwith probiotic lactic acid bacteriardquo LWT- Food Science andTechnology vol 76 pp 209ndash215 2017

[17] A Kumar and D Kumar ldquoCharacterization of Lactobacillusisolated from dairy samples for probiotic propertiesrdquoAnaerobevol 33 pp 117ndash123 2015

[18] P Li H Luo B Kong Q Liu and C Chen ldquoFormation of redmyoglobin derivatives and inhibition of spoilage bacteria in rawmeat batters by lactic acid bacteria and Staphylococcus xylosusrdquoLWT- Food Science and Technology vol 68 pp 251ndash257 2016

[19] Y Sha L Wang M Liu K Jiang F Xin and B Wang ldquoEffectsof lactic acid bacteria and the corresponding supernatant on thesurvival growth performance immune response and diseaseresistance of Litopenaeus vannameirdquo Aquaculture vol 452 pp28ndash36 2016

[20] M T Liong and N P Shah ldquoBile salt deconjugation ability bilesalt hydrolase activity and cholesterol co-precipitation ability oflactobacilli strainsrdquo International Dairy Journal vol 15 no 4pp 391ndash398 2005

[21] A C Ouwehand S Salminen and E Isolauri ldquoProbioticsan overview of beneficial effectsrdquo Antonie van Leeuwenhoek-Journal of Microbiology vol 82 no 1ndash4 pp 279ndash289 2002

[22] G Giraffa ldquoFunctionality of enterococci in dairy productsrdquoInternational Journal of Food Microbiology vol 88 no 2-3 pp215ndash222 2003

[23] AMercenier S Pavan and B Pot ldquoProbiotics as biotherapeuticagents Present knowledge and future prospectsrdquo Current Phar-maceutical Design vol 9 no 2 pp 175ndash191 2003

[24] V Strompfova and A Laukova ldquoIsolation and characterizationof faecal bifidobacteria and lactobacilli isolated from dogs andprimatesrdquo Anaerobe vol 29 pp 108ndash112 2014

[25] M Kargozari Z Emam-Djomeh H Gandomi R PartoviM Ghasemlou and I R Martin ldquoIdentification of selectedLactobacillus strains isolated from Siahmazgi cheese and studyon their behavior after inoculation in fermented-sausage modelmediumrdquo LWT- Food Science and Technology vol 62 no 2 pp1177ndash1183 2015

[26] M Nacef M Chevalier S Chollet D Drider and C FlahautldquoMALDI-TOFmass spectrometry for the identification of lacticacid bacteria isolated from a French cheese The MaroillesrdquoInternational Journal of Food Microbiology vol 247 pp 2ndash82017

[27] S Singh P Goswami R Singh and K J Heller ldquoApplication ofmolecular identification tools for Lactobacillus with a focus ondiscrimination between closely related species A reviewrdquo LWT-Food Science and Technology vol 42 no 2 pp 448ndash457 2009

[28] m Tarnberg t Jakobsson j Jonasson and u Forsum ldquoIdentifi-cation of randomly selected colonies of lactobacilli fromnormalvaginal fluid by pyrosequencing of the 16S rDNA variable V1and V3 regionsrdquo APMIS-Acta Pathologica Microbiologica etImmunologica Scandinavica vol 110 no 11 pp 802ndash810 2002

[29] J Yang Y Ji H Park et al ldquoSelection of functional lactic acidbacteria as starter cultures for the fermentation of Korean leek(Allium tuberosum Rottler ex Sprengel)rdquo International Journalof Food Microbiology vol 191 pp 164ndash171 2014

[30] M Carmen Collado and M Hernandez ldquoIdentification anddifferentiation of Lactobacillus Streptococcus and Bifidobac-terium species in fermented milk products with bifidobacteriardquoMicrobiological Research vol 162 no 1 pp 86ndash92 2007

[31] A M O Leite M A L Miguel R S Peixoto et al ldquoProbioticpotential of selected lactic acid bacteria strains isolated fromBrazilian kefir grainsrdquo Journal of Dairy Science vol 98 no 6pp 3622ndash3632 2015

[32] M M M Ahmed E E Hafez M A Mkamer H A-AElAbdelrrassoul and Y M A Mabrouk ldquoApplication of DNAtechnology to detect food infection with some pathogenicbacteria using 16S rDNA gene PCR-RFLP and sequencingrdquoJournal of Food Agriculture and Environment (JFAE) vol 12 no2 pp 202ndash206 2014

[33] D Bujnakova E Strakova and V Kmet ldquoInvitro evaluation ofthe safety and probiotic properties of Lactobacilli isolated fromchicken and calvesrdquo Anaerobe vol 29 pp 118ndash127 2014

[34] R Fuller ldquoProbiotics in man and animalsrdquo Journal of AppliedBacteriology vol 66 no 5 pp 365ndash378 1989

[35] J C de Man M Rogosa and M E Sharpe ldquoA medium for thecultivation of Lactobacillirdquo Journal of Applied Bacteriology vol23 no 1 pp 130ndash135 1960

[36] R R Ravula and N P Shah ldquoSelective enumeration of Lac-tobacillus casei from yogurts and fermented milk drinksrdquoBiotechnology Techniques vol 12 no 11 pp 819ndash822 1998


[37] B E Terzaghi and W E Sandine ldquoImproved medium forlactic streptococci and their bacteriophagesrdquo Journal of AppliedMicrobiology vol 29 no 6 pp 807ndash813 1975

[38] S Macwana and P M Muriana ldquoSpontaneous bacteriocinresistance in Listeria monocytogenes as a susceptibility screenfor identifying different mechanisms of resistance and modesof action by bacteriocins of lactic acid bacteriardquo Journal ofMicrobiological Methods vol 88 no 1 pp 7ndash13 2012

[39] I Javed Characterization of bacteriocin produced by lactic acidbacteria isolated from dairy products [PhD thesis] Quaid-i-Azam University Islamabad Pakistan 2009

[40] Y Sun X Lou X Zhu H Jiang and Q Gu ldquoIsolation and char-acterization of lactic acid bacteria producing bacteriocin fromnewborn infants fecesrdquo Journal of Bacteriology and Mycologyvol 1 no 2 pp 1ndash7 2014

[41] M Bosch J Nart S Audivert et al ldquoIsolation and characteriza-tion of probiotic strains for improving oral healthrdquo Archives ofOral Biolog vol 57 no 5 pp 539ndash549 2012

[42] O B Chahad M El Bour P Calo-Mata A Boudabous and JBarros-Velazquez ldquoDiscovery of novel biopreservation agentswith inhibitory effects on growth of food-borne pathogens andtheir application to seafood productsrdquoResearch inMicrobiologyvol 163 no 1 pp 44ndash54 2012

[43] S Schirru S D Todorov L Favaro et al ldquoSardinian goatrsquos milkas source of bacteriocinogenic potential protective culturesrdquoFood Control vol 25 no 1 pp 309ndash320 2012

[44] S V Hosseini S Arlindo K Bohme C Fernandez-No P Calo-Mata and J Barros-Velazquez ldquoMolecular and probiotic char-acterization of bacteriocin-producing Enterococcus faeciumstrains isolated from nonfermented animal foodsrdquo Journal ofApplied Microbiology vol 107 no 4 pp 1392ndash1403 2009

[45] Y A Chabbert ldquoLrsquoantibiogrammerdquo in Bacteriologie Medicale LLe Miror and M Veron Eds pp 205ndash212 Medecine ScienceParis France 1982

[46] B Haghshenas N Abdullah Y Nami D Radiah R Rosli andA Y Khosroushahi ldquoDifferent effects of two newly-isolatedprobiotic Lactobacillus plantarum 15HN and Lactococcus lactissubsp Lactis 44Lac strains from traditional dairy products oncancer cell linesrdquo Anaerobe vol 30 pp 51ndash59 2014

[47] R C Anderson A L Cookson W C McNabb W J Kelly andN C Roy ldquoLactobacillus plantarum DSM 2648 is a potentialprobiotic that enhances intestinal barrier functionrdquo FEMSMicrobiology Letters vol 309 no 2 pp 184ndash192 2010

[48] F Ait Ouali I Al Kassaa B Cudennec et al ldquoIdentificationof lactobacilli with inhibitory effect on biofilm formation bypathogenic bacteria on stainless steel surfacesrdquo InternationalJournal of Food Microbiology vol 191 pp 116ndash124 2014

[49] J L Balcazar I de Blas I Ruiz-Zarzuela D Vendrell OGirones and J L Muzquiz ldquoSequencing of variable regionsof the 16S rRNA gene for identification of lactic acid bacteriaisolated from the intestinal microbiota of healthy salmonidsrdquoComparative Immunology Microbiology amp Infectious Diseasesvol 30 no 2 pp 111ndash118 2007

[50] S Chakravorty S Sarkar andRGachhui ldquoIdentification of newconserved and variable regions in the 16S rRNA gene of aceticacid bacteria and acetobacteraceae familyrdquo Journal of MolecularBiology vol 49 no 5 pp 668ndash677 2015

[51] A Ahmadova S D Todorov Y Choiset et al ldquoEvaluation ofantimicrobial activity probiotic properties and safety of wildstrain Enterococcus faecium AQ71 isolated from AzerbaijaniMotal cheeserdquo Food Control vol 30 no 2 pp 631ndash641 2013

[52] SMathur and R Singh ldquoAntibiotic resistance in food lactic acidbacteriamdasha reviewrdquo International Journal of Food Microbiologyvol 105 no 3 pp 281ndash295 2005

[53] S Messaoudi A Madi H Prevost et al ldquoIn vitro evaluationof the probiotic potential of Lactobacillus salivarius SMXD51rdquoAnaerobe vol 18 no 6 pp 584ndash589 2012

[54] B A Neville and P Otoole ldquoProbiotic properties of Lacto-bacillus salivarius and closely related ctobacillus speciesrdquo FutureMicrobiology vol 5 no 5 pp 759ndash774 2010

[55] B Ladda T Theparee J Chimchang S Tanasupawat and MTaweechotipatr ldquoIn vitro modulation of tumor necrosis factor120572 production in THP-1 cells by lactic acid bacteria isolated fromhealthy human infantsrdquo Anaerobe vol 33 pp 109ndash116 2015

[56] M Ghanbari M Jami W Kneifel and K J Domig ldquoAntimi-crobial activity and partial characterization of bacteriocinsproduced by lactobacilli isolated from Sturgeon fishrdquo FoodControl vol 32 no 2 pp 379ndash385 2013

Stem Cells International

Hindawiwwwhindawicom Volume 2018


MEDIATORSINFLAMMATION

of

EndocrinologyInternational Journal of



Disease Markers


BioMed Research International

OncologyJournal of



Oxidative Medicine and Cellular Longevity


PPAR Research

Hindawi Publishing Corporation httpwwwhindawicom Volume 2013Hindawiwwwhindawicom

The Scientific World Journal

Volume 2018

Immunology ResearchHindawiwwwhindawicom Volume 2018

Journal of

ObesityJournal of



Computational and Mathematical Methods in Medicine


Behavioural Neurology

OphthalmologyJournal of


Diabetes ResearchJournal of



Research and TreatmentAIDS


Gastroenterology Research and Practice


Parkinsonrsquos Disease

Evidence-Based Complementary andAlternative Medicine

Volume 2018Hindawiwwwhindawicom

Submit your manuscripts atwwwhindawicom


species [25ndash29] Amplified ribosomal DNA restriction anal-ysis (ARDRA) and randomly amplified polymorphic DNA(RAPD) were successfully used in discriminating LAB [3031] The approaches include PCR-RFLP followed by directsequencing of the 16S rDNAgene and a BLAST search againstthe sequences of other organisms that are available at theNational Center for Biotechnology Information (NCBI) [32]

The isolation identification and characterization of novelLAB strains have two benefits The first is to reveal thecharacteristic taxonomy of the LAB and the second is toobtain promising beneficial and functional probiotic LAB[33 34] There is little research regarding the isolation andcharacterization of LAB from dairy products Therefore theaim of the present work was to isolate and to identify at themolecular level the lactic acid bacteria contained in rawmilkand fermented milk which were produced indigenously inSaudi Arabia These LAB were evaluated for their functionaltraits probiotic properties and ability to inhibit the growthof pathogenic and food poisoning bacteria

2 Materials and Methods

21 Samples andMedia Thirteen (05 kg) samples of rawmilkand traditional fermented milk from indigenous animalsprocured in Jeddah Province Saudi Arabia were used in thisstudy These samples were collected from the local marketand were stored in a fridge until use MRS agar and MRSbroth media (Oxoid119879119872 Thermo Fisher Scientific USA) wereused to isolate and support the growth of LAB and to inhibitthe growth of unwanted bacteria [35] MRS agar and brothwere also used to enumerate the LAB M17 agar and M17broth (Oxoid Thermo Fisher Scientific USA) were used forisolating the streptococci in dairy products [36 37] Bloodagar (Oxoid Thermo Fisher Scientific USA) and Muller-Hinton agar media (MHA) (HiMedia India) were used toevaluate hemolytic activity and antimicrobial activity of theLAB respectively

22 Isolation of LAB and Maintenance Method 50 g of thesamples was placed into sterile stomacher bags and then werediluted (110) with MRS broth processed to enrich the LABby stomaching sealed and incubated overnight at 30∘C [38]Lactic acid bacteria were isolated by 5-fold serial dilutionsAt the beginning 1mL of sample stomachate was added to9mL of sterile physiological water (085 NaCl) and wasfurther serially diluted Then 01mL aliquots of the samplesrsquosuitable dilutions were plated onto MRS agar and M17 agarTheplateswere incubated at 37∘C for 24-48 h in anaerobic jarsusing the AnaeroGen anaerobic system (Oxoid ThermoFisher Scientific USA) [25 39 40] and in microaerobicconditions (5 CO2) [41] MRS agar plates were used toenumerate the initial growth of the LAB in each sample [25]All experiments were performed in triplicate The resultingisolates were randomly selected from the medium surfaceand were streaked twice on fresh MRS to be purified [35]The isolated strains were maintained by culturing them inMRS broth medium at 30∘C and then storing at minus80∘C inMRS broth containing 50 (vv) glycerol MRS broth andM17 broth were used in all subsequent experiments [42 43]

Isolates from the stocks were subcultured in MRS broth forfurther studies

23 Phenotypic and Genotypic Identifications For all ofthe isolated strains the colony morphology on MRS solidmedium and M17 agar was determined visually and theirmotility was recorded using the hanging drop techniqueGram staining was performed to determine the cell mor-phology and Gram stain reaction of the isolates Cata-lase and cytochrome oxidase tests were also performed[39 42] As for genotypic identification cell pellets wereharvested from 2mL of overnight cultures (up to 2 times109 bacterial cells) of LAB grown in MRS broth by cen-trifugation for 10min at 5000 timesg and the supernatantwas discarded Then DNA extraction was done using aGeneJET Genomic DNA Purification Kit (cat no K0721Thermo Fisher Scientific Waltham MA USA) following themanufacturerrsquos instructions PCR was carried out and theuniversal primers were designed and synthesized (Medicbio-trade Germany) to amplify nearly the entire region of the16S rDNA gene the primers were 16SF27 (forward 51015840-AGAGTTTGATCCTGGCTCAG-31015840) and 16SR27 (reverse51015840-AAGGAGGTGATCCAGCCGCA-31015840) The primers weredesigned to amplify the 16S rDNA gene (sim16 kb [44])with GC contents of 50-60 for a higher melting temper-ature (Tm) to avoid nonspecific amplification [39] PCRconditions were one cycle of 5min of initial denaturationat 95∘C followed by 35 cycles of amplification Each cycleconsisted of denaturation at 95∘C for 30 sec annealing at58∘C for 30 sec and extension at 72∘C for 1min The lastcycle was 10min at 72∘C as a postextension step Elec-trophoresis in 05x TBE buffer was conducted on an 08agarose gel for the amplicons with a 1 kb DNA standard(TrackIt Invitrogen Waltham MA USA) and the gelswere stained with ethidium bromide and photographed Theamplicons were purified they were sequenced for the 16Sgene at Macrogen Inc (Seoul Korea) and they were BLAST-searched to detect similar sequences in the NCBI database(httpswwwncbinlmnihgov) using the Discovery StudioGene v15 (DSGene v15) program [32 42] A fragment ofsim1000 nt involving five variable (V) regions was further uti-lized Binary data metrics were entered into TFPGA (version13) and were analyzed using a qualitative route to generatea similarity coefficient The dissimilarity coefficients wereused to construct dendrograms by the sequential hierarchicaland nested clustering (neighbor joining (NJ)) method withNTSYSpc (version 210 Exeter Software)

24 Antibiotic Susceptibility Hemolytic Activity and AcidBile Tolerance The antibiotic susceptibility of the identi-fied LAB was determined by using the agar disk diffusionmethod [45] Fifteen antibiotics (Oxoid Thermo Fisher Sci-entific USA) were selected as representatives of the differentclasses of clinically important antibiotics Inoculates (106CFU mLminus1) of the LAB were swabbed onto the surfacesof Muller-Hinton agar plates and then the disks of antibi-otics (amoxicillin25120583g penicillin G10 units ceftriaxone30 120583g cefoxitin30120583g tobramycin10120583g oxacillin1120583g bac-itracin10 units chloramphenicol30120583g polymyxin B300









3 Results





Table1Genotypes

ofthes

elected46

isolated

LABas

16SrD

NAgene

sequ

encesa

lignm

entssubm

itted

totheN

CBIG

eneB

ankdatabase

Strain

no

Strain

name

GeneB

ankAc

cNo

Query

coverage

Maxident

Isolates

ourcea

2En

terococcus

faecium

Rashad3

KU324893

94

99

RCM

3En

terococcus

durans

Etim

ad1

KU324894

65

98

RCM

4En

terococcus

faecium

SMBM

3KU

324895

98

99

RCM

7La

ctobacillus

caseiM

SJ1

KU324896

98

99

CMCM

8La

ctobacillus

paracaseiN

MBM

1KU

324897

98

99

CMCM

9La

ctobacillus

caseiB

gShn

3KU

324898

99

99

CMCM

10La

ctobacillus

caseiM

asaLam

7KU

324899

95

99

CMCM

11La

ctobacillus

caseiD

wan5

KU324900

96

97

CMCM

12La

ctobacillus

plantarum

EyLan2

KU324901

89

97

CMCM

19La

ctobacillus

futsa

iiEM

BM2

KU324902

99

98

CCMCM

23Weis

sella

confusaSaEd

-7KU

324903

95

98

CCMCM

28En

terococcus

faecalisShbam40

KU324904

99

97

RGM

29En

terococcus

faecium

ZiNb3

KU324905

95

97

RGM

32En

terococcus

faecium

Gail-B

awZir8

KU324906

93

97

SYMCM

34En

terococcus

faecium

NSJ2

KU324907

99

96

SYMCM


ophilusB

inSlman8

KU324908

99

99

SYMCM

36La

ctococcuslactis

HadRa

mi9

KU324909

98

98

SYMCM

37En

terococcus

faecium

Marwh2

KU324910

97

99

QMCM

38En

terococcus

faecium

Mon

a3KU

324911

99

99

QMCM

39En

terococcus

faecium

SSJ3

KU324912

98

97

QMCM

40En

terococcus

lactisJedd

ah9

KU324913

97

98

QMCM


ophilusM

aNaL33

KU324914

97

98

QMCM

44En

terococcus

faeciumAd

eb3

KU324915

97

98

MMGM

45En

terococcus

faecium

ESJ4

KU324916

99

96

MMGM

46En

terococcus

durans

She7R

KU324917

96

97

MMGM

47La

ctobacillus

acidophilusH

adhram

aut4

KU324918

91

98

MMGM

50La

ctobacillus

acidophilusM

usallam2

KU324919

99

97

YMCM

53En

terococcus

faecium

BagH

om4

KU324920

99

99

RCAM


Omer9

KU324921

95

97

RCAM


Anw

r4KU

324922

77

96

SYMCA

M61


Zaki1

KU324923

98

98

SYMCA

M63


Salam7

KU324924

87

96

SYMCA

M68

Enterococcus

faecium

Ma7Fo

dKU

324925

98

98

SCMCM


JmaL3

KU324926

96

96

FCAM


Foad7

KU324927

95

96

FCAM

75Weis

sella

confusaAhM

d8KU

324928

99

99

BMCM

76Weis

sella

confusaTarim

4KU

324929

99

97

BMCM

77Weis

sella

confusaNoo

R1KU

324930

78

95

BMCM

78Weis

sella

confusaSaYu

n2KU

324931

98

98

BMCM

79Weis

sella

confusaMuK

alla5

KU324932

94

95

BMCM


Table1Con

tinued

Strain

no

Strain

name

GeneB

ankAc

cNo

Query

coverage

Maxident

Isolates

ourcea

80Weis

sella

confusaSYary1

KU324933

98

97

BMCM

81Weis

sella

confusaSho7ir

KU324934

96

98

BMCM

82Weis

sella

confusaFarag8

KU324935

97

97

BMCM

83Weis

sella

confusaA7G

afKU

324936

99

98

BMCM

92La

ctococcusgarvieaeZ

SJ5

KU324937

99

98

BMCM

93La

ctococcusgarvieaeE

mad4

KU324938

98

99

BMCM

a RCM

raw

cowmilk

RGMraw

goatmilk

RCA

Mraw

camelmilk

FCA

Mfrozencamelmilk

CMCM

cheesem

adefrom

cowmilk

CCM

CMcoo

kedcheese

madefrom

cowmilk

SCM

CMsalty

cheese

made

from

cowmilk

SYM

CMstirredyogu

rt(Laban)m

adefrom

cowmilk

SYM

CAMstirredyogu

rt(Laban)m

adefrom

camelmilk

YMCM

yogurtm

adefrom

cowmilk

BMCM

butterm

adefrom

cowmilk

QMCM

qeshta(cream

)madefrom

cowmilk

MMGMm

adheer

madefrom

goatmilk








Table2Th

eprofi

leof

theselected

LAB(n

=46

)for

mainprob

iotic

characteris

ticsa

ndtheantib

acteria

lactivity

spectraof

theirC

FSagainstseven

indicatoro

rganism

sAE

nterococcus

faecalisAT

CC29212B

EcoliAT

CC25922C

Salm

onellasppD

Shigella

sonn

eiAT

CC25931E

Staphylococcus

aureus

ATCC

25923F

MRS

AAT

CC43330

GL

isteria

monocytogenes

ATCC


n+

inhibitio

nzone

80ndash

100mm+

+inhibitio

nzone

101ndash

150mm+

++inh

ibition

zone

151ndash

200mm+

+++

inhibitio

nzone

200ndash300m

m

Strain

no-

code

Samples

ource

Strain

name

Accessionno

Hem

olysis

viabilityin

pH3medium

a

viabilityin

05

wv

bilea

Noof

antib

iotic

ssensitive

to15

AB

CD

EF

G

2RC

M2

Enterococcus

faecium

Rashad3

KU324893120574-hem

olysis

981plusmn0141

988plusmn0141

12+

++minus

+minus

++

3RC

M3

Enterococcus

durans

Etim

ad1

KU324894120574-hem

olysis

994plusmn0141

9775plusmn0354

12+

++minus

+minus

++

4RC

M4

Enterococcus

faecium

SMBM

3KU

324895120574-hem

olysis

991plusmn0283

981plusmn0283

13+

++minus

+minus

++

7CM

CM1

Lacto

bacilluscaseiMSJ1

KU324896120574-hem

olysis

9945plusmn0354

9905plusmn0212

9+

++++

+++

++

8CM

CM2

Lactobacillus

paracaseiN

MBM

1KU

324897120574-hem

olysis

9355plusmn0636

9845plusmn0212

10+

+++

+minus

+++

9CM

CM3

Lacto

bacilluscaseiBg

Shn3

KU324898120574-hem

olysis

9885plusmn0919

9925plusmn0212

9+

++

+minus

++

10CM

CM4

Lacto


7KU

324899120574-hem

olysis

975plusmn0283

974plusmn0424

9+

+++

+minus

+++

11CM

CM5

Lacto

bacilluscaseiDwan5

KU324900120574-hem

olysis

987plusmn0849

993plusmn0283

9+

+++

+++

++

12CM

CM6

Lactobacillus

plantarum

EyLan2

KU324901120574-hem

olysis

763plusmn0566

990plusmn0141

11+

++

++

++

19CC

MCM

1La

ctobacillus

futsa

iiEM

BM2

KU324902120574-hem

olysis

9835plusmn0778

918plusmn12

77

+++

+++minus

+++

23CC

MCM

5Weis

sella

confusaSaEd

-7KU

324903120572-hem

olysis

991plusmn0141

9545plusmn0354

10minus

minus+minusminus

+++

28RG

M5

Enterococcus

faecalisShbam40

KU324904120574-hem

olysis

822plusmn099

6745plusmn16

2612

minus++

++++

minusminus

+++

29RG

M6

Enterococcus

faecium

ZiNb3

KU324905120574-hem

olysis

994plusmn0141

996plusmn0141

12+

+minus

++

+++

32SY

MCM

3En

terococcus

faecium

Gail-B

awZir8

KU324906120574-hem

olysis

991plusmn0283

741plusmn2263

12++

+++

+++

++

34SY

MCM

5En

terococcus

faecium

NSJ2

KU324907120574-hem

olysis

9896plusmn0078

976plusmn0424

11+

++

++minus

+++

++35

SYMCM

6Streptococcustherm

ophilusB

inSlman8

KU324908120572-hem

olysis

6735plusmn14

859565plusmn0636

11minus

++minus

++++

minus+

36SY

MCM

7La

ctococcuslactis

HadRa

mi9

KU324909120574-hem

olysis

9895plusmn0212

655plusmn19

812

minus+

+++

minusminus

++37

QMCM

1En

terococcus

faecium

Marwh2

KU324910120574-hem

olysis

985plusmn0424

8365plusmn12

0212

++minus

+minus

++

38QMCM

2En

terococcus

faecium

Mon

a3KU

324911120574-hem

olysis

9755plusmn0354

6815plusmn16

2611

+++

++minus

+++

39QMCM

3En

terococcus

faecium

SSJ3

KU324912120574-hem

olysis

983plusmn0283

609plusmn12

7312

+++minus

+minus

++

40QMCM

4En

terococcus

lactisJedd

ah9

KU324913120574-hem

olysis

9715plusmn0354

6715plusmn0636

11minus

++

+minus

++

42QMC M

6Streptococcustherm

ophilusM

aNaL33

KU324914120572-hem

olysis

6085plusmn14

85951plusmn0424

11minus

++minus

++++

minus+

44MMGM1

Enterococcus

faecium

Adeb3

KU324915120574-hem

olysis

985plusmn0566

9735plusmn0636

8+

+++

++minus

minus++

45MMGM2

Enterococcus

faecium

ESJ4

KU324916120574-hem

olysis

991plusmn0283

598plusmn0283

12++

+minus

+minus

+++

46MMGM3

Enterococcus

durans

She7R

KU324917120574-hem

olysis

956plusmn0424

667plusmn12

7311

+++minus

+minus

+++

47MMGM4

Lactobacillus

acidophilusH

adhram

aut4

KU324918120574-hem

olysis

8265plusmn0636

907plusmn1131

10minus

+++

++minus

+++

50YM

CM3

Lactobacillus

acidophilusM

usallam2

KU324919120574-hem

olysis

9225plusmn0495

9945plusmn0212

11minus

+++

++minus

+++

+53

RCAM1

Enterococcus

faecium

BagH

om4

KU324920120574-hem

olysis

995plusmn01 41

9825plusmn0495

13+

++minus

+minus

++

58RC

AM6


Omer9

KU324921120574-hem

olysis

6455plusmn2758

975plusmn0283

13minus

++

++minus

minus++

60SY

MCA

M2


Anw

r4KU

324922120574-hem

olysis

7285plusmn12

029745plusmn0495

14minus

++

++minus

+++

++61

SYMCA

M3


Zaki1

KU324923120574-hem

olysis

7575plusmn19

09971plusmn0849

14minus

++

++minus

minus+

63SY

MCA

M5


Salam7

KU324924120574-hem

olysis

7445plusmn0636

9895plusmn0212

14minus

++

++minus

+++

+68

SCMCM

4En

terococcus

faecium

Ma7Fo

dKU

324925120574-hem

olysis

9845plusmn0636

377plusmn0990

13+

+++

++minus

++

73FC

AM4


JmaL3

KU324926120574-hem

olysis

5285plusmn0636

9555plusmn0636

13minus

+minusminus

++

++74

FCAM5


Foad7

KU324927120574-hem

o lysis

5675plusmn14

85953plusmn0566

12minus

+minusminus

++

++75

BMCM

1Weis

sella

confusaAhM

d8KU

324928120572-hem

olysis

9945plusmn0212

953plusmn0424

10minus

minus+minusminus

minusminus

76BM

CM2

Weis

sella

confusaTarim

4KU

324929120572-hem

olysis

9915plusmn0212

952plusmn0424

10minus

minus+minusminus

++++minus

77BM

CM3

Weis

sella

confusaNoo

R1KU

324930120572-hem

olysis

9955plusmn0071

9655plusmn0778

10++

++minusminus

+++minus

78BM

CM4

Weis

sella

confusaSaYu

n2KU

324931120572-hem

olysis

9945plusmn071

9635plusmn0495

10minus

++++minusminus

minusminus

79BM

CM5

Weis

sella

confusaMuK

alla5

KU324932120572-hem

olysis

709plusmn0707

9595plusmn0212

11minus

+minusminus

+++

+


Table2Con

tinued

Strain

no-

code

Samples

ource

Strain

name

Accessionno

Hem

olysis

viabilityin

pH3medium

a

viabilityin

05

wv

bilea

Noof

antib

iotic

ssensitive

to15

AB

CD

EF

G

80BM

CM6

Weis

sella

confusaSYary1

KU324933

120572-hem

olysis

992plusmn0141

982plusmn0424

11minus


+81

BMCM

7Weis

sella

confusaSho7ir

KU324934

120572-hem

olysis

969plusmn0566

949plusmn0849

10++

minus+

++

++

82BM

CM8

Weis

sella

confusaFarag8

KU324935

120572-hem

olysis

973plusmn0424

9565plusmn0636

10++


83BM

CM9

Weis

sella

confusaA7G

afKU

324936

120572-hem

olysis

9675plusmn0212

9515plusmn0636

10++

minus+minusminus

+minus

92BM

CM18


SJ5

KU324937

120572-hem

olysis

885plusmn0707

9865plusmn0354

11minus


93BM

CM19


mad4

KU324938

120572-hem

olysis

860plusmn0707

992plusmn0141

11minus


a Resultsarep

resented

asthem

eanvalueo

fdup

licatetria

lsplusmnstandard

deviation(SD)


3770 50-70 701-899 90-95 951-979 98-9955



0

5

10

15

20

25

Num

ber o

f str

ains





4 Discussion









5 Conclusions




Acknowledgments


References
































































of




Disease Markers



OncologyJournal of





PPAR Research



Volume 2018


Journal of

ObesityJournal of



























3 Results





Table1Genotypes

ofthes

elected46

isolated

LABas

16SrD

NAgene

sequ

encesa

lignm

entssubm

itted

totheN

CBIG

eneB

ankdatabase

Strain

no

Strain

name

GeneB

ankAc

cNo

Query

coverage

Maxident

Isolates

ourcea

2En

terococcus

faecium

Rashad3

KU324893

94

99

RCM

3En

terococcus

durans

Etim

ad1

KU324894

65

98

RCM

4En

terococcus

faecium

SMBM

3KU

324895

98

99

RCM

7La

ctobacillus

caseiM

SJ1

KU324896

98

99

CMCM

8La

ctobacillus

paracaseiN

MBM

1KU

324897

98

99

CMCM

9La

ctobacillus

caseiB

gShn

3KU

324898

99

99

CMCM

10La

ctobacillus

caseiM

asaLam

7KU

324899

95

99

CMCM

11La

ctobacillus

caseiD

wan5

KU324900

96

97

CMCM

12La

ctobacillus

plantarum

EyLan2

KU324901

89

97

CMCM

19La

ctobacillus

futsa

iiEM

BM2

KU324902

99

98

CCMCM

23Weis

sella

confusaSaEd

-7KU

324903

95

98

CCMCM

28En

terococcus

faecalisShbam40

KU324904

99

97

RGM

29En

terococcus

faecium

ZiNb3

KU324905

95

97

RGM

32En

terococcus

faecium

Gail-B

awZir8

KU324906

93

97

SYMCM

34En

terococcus

faecium

NSJ2

KU324907

99

96

SYMCM


ophilusB

inSlman8

KU324908

99

99

SYMCM

36La

ctococcuslactis

HadRa

mi9

KU324909

98

98

SYMCM

37En

terococcus

faecium

Marwh2

KU324910

97

99

QMCM

38En

terococcus

faecium

Mon

a3KU

324911

99

99

QMCM

39En

terococcus

faecium

SSJ3

KU324912

98

97

QMCM

40En

terococcus

lactisJedd

ah9

KU324913

97

98

QMCM


ophilusM

aNaL33

KU324914

97

98

QMCM

44En

terococcus

faeciumAd

eb3

KU324915

97

98

MMGM

45En

terococcus

faecium

ESJ4

KU324916

99

96

MMGM

46En

terococcus

durans

She7R

KU324917

96

97

MMGM

47La

ctobacillus

acidophilusH

adhram

aut4

KU324918

91

98

MMGM

50La

ctobacillus

acidophilusM

usallam2

KU324919

99

97

YMCM

53En

terococcus

faecium

BagH

om4

KU324920

99

99

RCAM


Omer9

KU324921

95

97

RCAM


Anw

r4KU

324922

77

96

SYMCA

M61


Zaki1

KU324923

98

98

SYMCA

M63


Salam7

KU324924

87

96

SYMCA

M68

Enterococcus

faecium

Ma7Fo

dKU

324925

98

98

SCMCM


JmaL3

KU324926

96

96

FCAM


Foad7

KU324927

95

96

FCAM

75Weis

sella

confusaAhM

d8KU

324928

99

99

BMCM

76Weis

sella

confusaTarim

4KU

324929

99

97

BMCM

77Weis

sella

confusaNoo

R1KU

324930

78

95

BMCM

78Weis

sella

confusaSaYu

n2KU

324931

98

98

BMCM

79Weis

sella

confusaMuK

alla5

KU324932

94

95

BMCM


Table1Con

tinued

Strain

no

Strain

name

GeneB

ankAc

cNo

Query

coverage

Maxident

Isolates

ourcea

80Weis

sella

confusaSYary1

KU324933

98

97

BMCM

81Weis

sella

confusaSho7ir

KU324934

96

98

BMCM

82Weis

sella

confusaFarag8

KU324935

97

97

BMCM

83Weis

sella

confusaA7G

afKU

324936

99

98

BMCM

92La

ctococcusgarvieaeZ

SJ5

KU324937

99

98

BMCM

93La

ctococcusgarvieaeE

mad4

KU324938

98

99

BMCM

a RCM

raw

cowmilk

RGMraw

goatmilk

RCA

Mraw

camelmilk

FCA

Mfrozencamelmilk

CMCM

cheesem

adefrom

cowmilk

CCM

CMcoo

kedcheese

madefrom

cowmilk

SCM

CMsalty

cheese

made

from

cowmilk

SYM

CMstirredyogu

rt(Laban)m

adefrom

cowmilk

SYM

CAMstirredyogu

rt(Laban)m

adefrom

camelmilk

YMCM

yogurtm

adefrom

cowmilk

BMCM

butterm

adefrom

cowmilk

QMCM

qeshta(cream

)madefrom

cowmilk

MMGMm

adheer

madefrom

goatmilk








Table2Th

eprofi

leof

theselected

LAB(n

=46

)for

mainprob

iotic

characteris

ticsa

ndtheantib

acteria

lactivity

spectraof

theirC

FSagainstseven

indicatoro

rganism

sAE

nterococcus

faecalisAT

CC29212B

EcoliAT

CC25922C

Salm

onellasppD

Shigella

sonn

eiAT

CC25931E

Staphylococcus

aureus

ATCC

25923F

MRS

AAT

CC43330

GL

isteria

monocytogenes

ATCC


n+

inhibitio

nzone

80ndash

100mm+

+inhibitio

nzone

101ndash

150mm+

++inh

ibition

zone

151ndash

200mm+

+++

inhibitio

nzone

200ndash300m

m

Strain

no-

code

Samples

ource

Strain

name

Accessionno

Hem

olysis

viabilityin

pH3medium

a

viabilityin

05

wv

bilea

Noof

antib

iotic

ssensitive

to15

AB

CD

EF

G

2RC

M2

Enterococcus

faecium

Rashad3

KU324893120574-hem

olysis

981plusmn0141

988plusmn0141

12+

++minus

+minus

++

3RC

M3

Enterococcus

durans

Etim

ad1

KU324894120574-hem

olysis

994plusmn0141

9775plusmn0354

12+

++minus

+minus

++

4RC

M4

Enterococcus

faecium

SMBM

3KU

324895120574-hem

olysis

991plusmn0283

981plusmn0283

13+

++minus

+minus

++

7CM

CM1

Lacto

bacilluscaseiMSJ1

KU324896120574-hem

olysis

9945plusmn0354

9905plusmn0212

9+

++++

+++

++

8CM

CM2

Lactobacillus

paracaseiN

MBM

1KU

324897120574-hem

olysis

9355plusmn0636

9845plusmn0212

10+

+++

+minus

+++

9CM

CM3

Lacto

bacilluscaseiBg

Shn3

KU324898120574-hem

olysis

9885plusmn0919

9925plusmn0212

9+

++

+minus

++

10CM

CM4

Lacto


7KU

324899120574-hem

olysis

975plusmn0283

974plusmn0424

9+

+++

+minus

+++

11CM

CM5

Lacto

bacilluscaseiDwan5

KU324900120574-hem

olysis

987plusmn0849

993plusmn0283

9+

+++

+++

++

12CM

CM6

Lactobacillus

plantarum

EyLan2

KU324901120574-hem

olysis

763plusmn0566

990plusmn0141

11+

++

++

++

19CC

MCM

1La

ctobacillus

futsa

iiEM

BM2

KU324902120574-hem

olysis

9835plusmn0778

918plusmn12

77

+++

+++minus

+++

23CC

MCM

5Weis

sella

confusaSaEd

-7KU

324903120572-hem

olysis

991plusmn0141

9545plusmn0354

10minus

minus+minusminus

+++

28RG

M5

Enterococcus

faecalisShbam40

KU324904120574-hem

olysis

822plusmn099

6745plusmn16

2612

minus++

++++

minusminus

+++

29RG

M6

Enterococcus

faecium

ZiNb3

KU324905120574-hem

olysis

994plusmn0141

996plusmn0141

12+

+minus

++

+++

32SY

MCM

3En

terococcus

faecium

Gail-B

awZir8

KU324906120574-hem

olysis

991plusmn0283

741plusmn2263

12++

+++

+++

++

34SY

MCM

5En

terococcus

faecium

NSJ2

KU324907120574-hem

olysis

9896plusmn0078

976plusmn0424

11+

++

++minus

+++

++35

SYMCM

6Streptococcustherm

ophilusB

inSlman8

KU324908120572-hem

olysis

6735plusmn14

859565plusmn0636

11minus

++minus

++++

minus+

36SY

MCM

7La

ctococcuslactis

HadRa

mi9

KU324909120574-hem

olysis

9895plusmn0212

655plusmn19

812

minus+

+++

minusminus

++37

QMCM

1En

terococcus

faecium

Marwh2

KU324910120574-hem

olysis

985plusmn0424

8365plusmn12

0212

++minus

+minus

++

38QMCM

2En

terococcus

faecium

Mon

a3KU

324911120574-hem

olysis

9755plusmn0354

6815plusmn16

2611

+++

++minus

+++

39QMCM

3En

terococcus

faecium

SSJ3

KU324912120574-hem

olysis

983plusmn0283

609plusmn12

7312

+++minus

+minus

++

40QMCM

4En

terococcus

lactisJedd

ah9

KU324913120574-hem

olysis

9715plusmn0354

6715plusmn0636

11minus

++

+minus

++

42QMC M

6Streptococcustherm

ophilusM

aNaL33

KU324914120572-hem

olysis

6085plusmn14

85951plusmn0424

11minus

++minus

++++

minus+

44MMGM1

Enterococcus

faecium

Adeb3

KU324915120574-hem

olysis

985plusmn0566

9735plusmn0636

8+

+++

++minus

minus++

45MMGM2

Enterococcus

faecium

ESJ4

KU324916120574-hem

olysis

991plusmn0283

598plusmn0283

12++

+minus

+minus

+++

46MMGM3

Enterococcus

durans

She7R

KU324917120574-hem

olysis

956plusmn0424

667plusmn12

7311

+++minus

+minus

+++

47MMGM4

Lactobacillus

acidophilusH

adhram

aut4

KU324918120574-hem

olysis

8265plusmn0636

907plusmn1131

10minus

+++

++minus

+++

50YM

CM3

Lactobacillus

acidophilusM

usallam2

KU324919120574-hem

olysis

9225plusmn0495

9945plusmn0212

11minus

+++

++minus

+++

+53

RCAM1

Enterococcus

faecium

BagH

om4

KU324920120574-hem

olysis

995plusmn01 41

9825plusmn0495

13+

++minus

+minus

++

58RC

AM6


Omer9

KU324921120574-hem

olysis

6455plusmn2758

975plusmn0283

13minus

++

++minus

minus++

60SY

MCA

M2


Anw

r4KU

324922120574-hem

olysis

7285plusmn12

029745plusmn0495

14minus

++

++minus

+++

++61

SYMCA

M3


Zaki1

KU324923120574-hem

olysis

7575plusmn19

09971plusmn0849

14minus

++

++minus

minus+

63SY

MCA

M5


Salam7

KU324924120574-hem

olysis

7445plusmn0636

9895plusmn0212

14minus

++

++minus

+++

+68

SCMCM

4En

terococcus

faecium

Ma7Fo

dKU

324925120574-hem

olysis

9845plusmn0636

377plusmn0990

13+

+++

++minus

++

73FC

AM4


JmaL3

KU324926120574-hem

olysis

5285plusmn0636

9555plusmn0636

13minus

+minusminus

++

++74

FCAM5


Foad7

KU324927120574-hem

o lysis

5675plusmn14

85953plusmn0566

12minus

+minusminus

++

++75

BMCM

1Weis

sella

confusaAhM

d8KU

324928120572-hem

olysis

9945plusmn0212

953plusmn0424

10minus

minus+minusminus

minusminus

76BM

CM2

Weis

sella

confusaTarim

4KU

324929120572-hem

olysis

9915plusmn0212

952plusmn0424

10minus

minus+minusminus

++++minus

77BM

CM3

Weis

sella

confusaNoo

R1KU

324930120572-hem

olysis

9955plusmn0071

9655plusmn0778

10++

++minusminus

+++minus

78BM

CM4

Weis

sella

confusaSaYu

n2KU

324931120572-hem

olysis

9945plusmn071

9635plusmn0495

10minus

++++minusminus

minusminus

79BM

CM5

Weis

sella

confusaMuK

alla5

KU324932120572-hem

olysis

709plusmn0707

9595plusmn0212

11minus

+minusminus

+++

+


Table2Con

tinued

Strain

no-

code

Samples

ource

Strain

name

Accessionno

Hem

olysis

viabilityin

pH3medium

a

viabilityin

05

wv

bilea

Noof

antib

iotic

ssensitive

to15

AB

CD

EF

G

80BM

CM6

Weis

sella

confusaSYary1

KU324933

120572-hem

olysis

992plusmn0141

982plusmn0424

11minus


+81

BMCM

7Weis

sella

confusaSho7ir

KU324934

120572-hem

olysis

969plusmn0566

949plusmn0849

10++

minus+

++

++

82BM

CM8

Weis

sella

confusaFarag8

KU324935

120572-hem

olysis

973plusmn0424

9565plusmn0636

10++


83BM

CM9

Weis

sella

confusaA7G

afKU

324936

120572-hem

olysis

9675plusmn0212

9515plusmn0636

10++

minus+minusminus

+minus

92BM

CM18


SJ5

KU324937

120572-hem

olysis

885plusmn0707

9865plusmn0354

11minus


93BM

CM19


mad4

KU324938

120572-hem

olysis

860plusmn0707

992plusmn0141

11minus


a Resultsarep

resented

asthem

eanvalueo

fdup

licatetria

lsplusmnstandard

deviation(SD)


3770 50-70 701-899 90-95 951-979 98-9955



0

5

10

15

20

25

Num

ber o

f str

ains





4 Discussion









5 Conclusions




Acknowledgments


References
































































of




Disease Markers



OncologyJournal of





PPAR Research



Volume 2018


Journal of

ObesityJournal of




















Table1Genotypes

ofthes

elected46

isolated

LABas

16SrD

NAgene

sequ

encesa

lignm

entssubm

itted

totheN

CBIG

eneB

ankdatabase

Strain

no

Strain

name

GeneB

ankAc

cNo

Query

coverage

Maxident

Isolates

ourcea

2En

terococcus

faecium

Rashad3

KU324893

94

99

RCM

3En

terococcus

durans

Etim

ad1

KU324894

65

98

RCM

4En

terococcus

faecium

SMBM

3KU

324895

98

99

RCM

7La

ctobacillus

caseiM

SJ1

KU324896

98

99

CMCM

8La

ctobacillus

paracaseiN

MBM

1KU

324897

98

99

CMCM

9La

ctobacillus

caseiB

gShn

3KU

324898

99

99

CMCM

10La

ctobacillus

caseiM

asaLam

7KU

324899

95

99

CMCM

11La

ctobacillus

caseiD

wan5

KU324900

96

97

CMCM

12La

ctobacillus

plantarum

EyLan2

KU324901

89

97

CMCM

19La

ctobacillus

futsa

iiEM

BM2

KU324902

99

98

CCMCM

23Weis

sella

confusaSaEd

-7KU

324903

95

98

CCMCM

28En

terococcus

faecalisShbam40

KU324904

99

97

RGM

29En

terococcus

faecium

ZiNb3

KU324905

95

97

RGM

32En

terococcus

faecium

Gail-B

awZir8

KU324906

93

97

SYMCM

34En

terococcus

faecium

NSJ2

KU324907

99

96

SYMCM


ophilusB

inSlman8

KU324908

99

99

SYMCM

36La

ctococcuslactis

HadRa

mi9

KU324909

98

98

SYMCM

37En

terococcus

faecium

Marwh2

KU324910

97

99

QMCM

38En

terococcus

faecium

Mon

a3KU

324911

99

99

QMCM

39En

terococcus

faecium

SSJ3

KU324912

98

97

QMCM

40En

terococcus

lactisJedd

ah9

KU324913

97

98

QMCM


ophilusM

aNaL33

KU324914

97

98

QMCM

44En

terococcus

faeciumAd

eb3

KU324915

97

98

MMGM

45En

terococcus

faecium

ESJ4

KU324916

99

96

MMGM

46En

terococcus

durans

She7R

KU324917

96

97

MMGM

47La

ctobacillus

acidophilusH

adhram

aut4

KU324918

91

98

MMGM

50La

ctobacillus

acidophilusM

usallam2

KU324919

99

97

YMCM

53En

terococcus

faecium

BagH

om4

KU324920

99

99

RCAM


Omer9

KU324921

95

97

RCAM


Anw

r4KU

324922

77

96

SYMCA

M61


Zaki1

KU324923

98

98

SYMCA

M63


Salam7

KU324924

87

96

SYMCA

M68

Enterococcus

faecium

Ma7Fo

dKU

324925

98

98

SCMCM


JmaL3

KU324926

96

96

FCAM


Foad7

KU324927

95

96

FCAM

75Weis

sella

confusaAhM

d8KU

324928

99

99

BMCM

76Weis

sella

confusaTarim

4KU

324929

99

97

BMCM

77Weis

sella

confusaNoo

R1KU

324930

78

95

BMCM

78Weis

sella

confusaSaYu

n2KU

324931

98

98

BMCM

79Weis

sella

confusaMuK

alla5

KU324932

94

95

BMCM


Table1Con

tinued

Strain

no

Strain

name

GeneB

ankAc

cNo

Query

coverage

Maxident

Isolates

ourcea

80Weis

sella

confusaSYary1

KU324933

98

97

BMCM

81Weis

sella

confusaSho7ir

KU324934

96

98

BMCM

82Weis

sella

confusaFarag8

KU324935

97

97

BMCM

83Weis

sella

confusaA7G

afKU

324936

99

98

BMCM

92La

ctococcusgarvieaeZ

SJ5

KU324937

99

98

BMCM

93La

ctococcusgarvieaeE

mad4

KU324938

98

99

BMCM

a RCM

raw

cowmilk

RGMraw

goatmilk

RCA

Mraw

camelmilk

FCA

Mfrozencamelmilk

CMCM

cheesem

adefrom

cowmilk

CCM

CMcoo

kedcheese

madefrom

cowmilk

SCM

CMsalty

cheese

made

from

cowmilk

SYM

CMstirredyogu

rt(Laban)m

adefrom

cowmilk

SYM

CAMstirredyogu

rt(Laban)m

adefrom

camelmilk

YMCM

yogurtm

adefrom

cowmilk

BMCM

butterm

adefrom

cowmilk

QMCM

qeshta(cream

)madefrom

cowmilk

MMGMm

adheer

madefrom

goatmilk








Table2Th

eprofi

leof

theselected

LAB(n

=46

)for

mainprob

iotic

characteris

ticsa

ndtheantib

acteria

lactivity

spectraof

theirC

FSagainstseven

indicatoro

rganism

sAE

nterococcus

faecalisAT

CC29212B

EcoliAT

CC25922C

Salm

onellasppD

Shigella

sonn

eiAT

CC25931E

Staphylococcus

aureus

ATCC

25923F

MRS

AAT

CC43330

GL

isteria

monocytogenes

ATCC


n+

inhibitio

nzone

80ndash

100mm+

+inhibitio

nzone

101ndash

150mm+

++inh

ibition

zone

151ndash

200mm+

+++

inhibitio

nzone

200ndash300m

m

Strain

no-

code

Samples

ource

Strain

name

Accessionno

Hem

olysis

viabilityin

pH3medium

a

viabilityin

05

wv

bilea

Noof

antib

iotic

ssensitive

to15

AB

CD

EF

G

2RC

M2

Enterococcus

faecium

Rashad3

KU324893120574-hem

olysis

981plusmn0141

988plusmn0141

12+

++minus

+minus

++

3RC

M3

Enterococcus

durans

Etim

ad1

KU324894120574-hem

olysis

994plusmn0141

9775plusmn0354

12+

++minus

+minus

++

4RC

M4

Enterococcus

faecium

SMBM

3KU

324895120574-hem

olysis

991plusmn0283

981plusmn0283

13+

++minus

+minus

++

7CM

CM1

Lacto

bacilluscaseiMSJ1

KU324896120574-hem

olysis

9945plusmn0354

9905plusmn0212

9+

++++

+++

++

8CM

CM2

Lactobacillus

paracaseiN

MBM

1KU

324897120574-hem

olysis

9355plusmn0636

9845plusmn0212

10+

+++

+minus

+++

9CM

CM3

Lacto

bacilluscaseiBg

Shn3

KU324898120574-hem

olysis

9885plusmn0919

9925plusmn0212

9+

++

+minus

++

10CM

CM4

Lacto


7KU

324899120574-hem

olysis

975plusmn0283

974plusmn0424

9+

+++

+minus

+++

11CM

CM5

Lacto

bacilluscaseiDwan5

KU324900120574-hem

olysis

987plusmn0849

993plusmn0283

9+

+++

+++

++

12CM

CM6

Lactobacillus

plantarum

EyLan2

KU324901120574-hem

olysis

763plusmn0566

990plusmn0141

11+

++

++

++

19CC

MCM

1La

ctobacillus

futsa

iiEM

BM2

KU324902120574-hem

olysis

9835plusmn0778

918plusmn12

77

+++

+++minus

+++

23CC

MCM

5Weis

sella

confusaSaEd

-7KU

324903120572-hem

olysis

991plusmn0141

9545plusmn0354

10minus

minus+minusminus

+++

28RG

M5

Enterococcus

faecalisShbam40

KU324904120574-hem

olysis

822plusmn099

6745plusmn16

2612

minus++

++++

minusminus

+++

29RG

M6

Enterococcus

faecium

ZiNb3

KU324905120574-hem

olysis

994plusmn0141

996plusmn0141

12+

+minus

++

+++

32SY

MCM

3En

terococcus

faecium

Gail-B

awZir8

KU324906120574-hem

olysis

991plusmn0283

741plusmn2263

12++

+++

+++

++

34SY

MCM

5En

terococcus

faecium

NSJ2

KU324907120574-hem

olysis

9896plusmn0078

976plusmn0424

11+

++

++minus

+++

++35

SYMCM

6Streptococcustherm

ophilusB

inSlman8

KU324908120572-hem

olysis

6735plusmn14

859565plusmn0636

11minus

++minus

++++

minus+

36SY

MCM

7La

ctococcuslactis

HadRa

mi9

KU324909120574-hem

olysis

9895plusmn0212

655plusmn19

812

minus+

+++

minusminus

++37

QMCM

1En

terococcus

faecium

Marwh2

KU324910120574-hem

olysis

985plusmn0424

8365plusmn12

0212

++minus

+minus

++

38QMCM

2En

terococcus

faecium

Mon

a3KU

324911120574-hem

olysis

9755plusmn0354

6815plusmn16

2611

+++

++minus

+++

39QMCM

3En

terococcus

faecium

SSJ3

KU324912120574-hem

olysis

983plusmn0283

609plusmn12

7312

+++minus

+minus

++

40QMCM

4En

terococcus

lactisJedd

ah9

KU324913120574-hem

olysis

9715plusmn0354

6715plusmn0636

11minus

++

+minus

++

42QMC M

6Streptococcustherm

ophilusM

aNaL33

KU324914120572-hem

olysis

6085plusmn14

85951plusmn0424

11minus

++minus

++++

minus+

44MMGM1

Enterococcus

faecium

Adeb3

KU324915120574-hem

olysis

985plusmn0566

9735plusmn0636

8+

+++

++minus

minus++

45MMGM2

Enterococcus

faecium

ESJ4

KU324916120574-hem

olysis

991plusmn0283

598plusmn0283

12++

+minus

+minus

+++

46MMGM3

Enterococcus

durans

She7R

KU324917120574-hem

olysis

956plusmn0424

667plusmn12

7311

+++minus

+minus

+++

47MMGM4

Lactobacillus

acidophilusH

adhram

aut4

KU324918120574-hem

olysis

8265plusmn0636

907plusmn1131

10minus

+++

++minus

+++

50YM

CM3

Lactobacillus

acidophilusM

usallam2

KU324919120574-hem

olysis

9225plusmn0495

9945plusmn0212

11minus

+++

++minus

+++

+53

RCAM1

Enterococcus

faecium

BagH

om4

KU324920120574-hem

olysis

995plusmn01 41

9825plusmn0495

13+

++minus

+minus

++

58RC

AM6


Omer9

KU324921120574-hem

olysis

6455plusmn2758

975plusmn0283

13minus

++

++minus

minus++

60SY

MCA

M2


Anw

r4KU

324922120574-hem

olysis

7285plusmn12

029745plusmn0495

14minus

++

++minus

+++

++61

SYMCA

M3


Zaki1

KU324923120574-hem

olysis

7575plusmn19

09971plusmn0849

14minus

++

++minus

minus+

63SY

MCA

M5


Salam7

KU324924120574-hem

olysis

7445plusmn0636

9895plusmn0212

14minus

++

++minus

+++

+68

SCMCM

4En

terococcus

faecium

Ma7Fo

dKU

324925120574-hem

olysis

9845plusmn0636

377plusmn0990

13+

+++

++minus

++

73FC

AM4


JmaL3

KU324926120574-hem

olysis

5285plusmn0636

9555plusmn0636

13minus

+minusminus

++

++74

FCAM5


Foad7

KU324927120574-hem

o lysis

5675plusmn14

85953plusmn0566

12minus

+minusminus

++

++75

BMCM

1Weis

sella

confusaAhM

d8KU

324928120572-hem

olysis

9945plusmn0212

953plusmn0424

10minus

minus+minusminus

minusminus

76BM

CM2

Weis

sella

confusaTarim

4KU

324929120572-hem

olysis

9915plusmn0212

952plusmn0424

10minus

minus+minusminus

++++minus

77BM

CM3

Weis

sella

confusaNoo

R1KU

324930120572-hem

olysis

9955plusmn0071

9655plusmn0778

10++

++minusminus

+++minus

78BM

CM4

Weis

sella

confusaSaYu

n2KU

324931120572-hem

olysis

9945plusmn071

9635plusmn0495

10minus

++++minusminus

minusminus

79BM

CM5

Weis

sella

confusaMuK

alla5

KU324932120572-hem

olysis

709plusmn0707

9595plusmn0212

11minus

+minusminus

+++

+


Table2Con

tinued

Strain

no-

code

Samples

ource

Strain

name

Accessionno

Hem

olysis

viabilityin

pH3medium

a

viabilityin

05

wv

bilea

Noof

antib

iotic

ssensitive

to15

AB

CD

EF

G

80BM

CM6

Weis

sella

confusaSYary1

KU324933

120572-hem

olysis

992plusmn0141

982plusmn0424

11minus


+81

BMCM

7Weis

sella

confusaSho7ir

KU324934

120572-hem

olysis

969plusmn0566

949plusmn0849

10++

minus+

++

++

82BM

CM8

Weis

sella

confusaFarag8

KU324935

120572-hem

olysis

973plusmn0424

9565plusmn0636

10++


83BM

CM9

Weis

sella

confusaA7G

afKU

324936

120572-hem

olysis

9675plusmn0212

9515plusmn0636

10++

minus+minusminus

+minus

92BM

CM18


SJ5

KU324937

120572-hem

olysis

885plusmn0707

9865plusmn0354

11minus


93BM

CM19


mad4

KU324938

120572-hem

olysis

860plusmn0707

992plusmn0141

11minus


a Resultsarep

resented

asthem

eanvalueo

fdup

licatetria

lsplusmnstandard

deviation(SD)


3770 50-70 701-899 90-95 951-979 98-9955



0

5

10

15

20

25

Num

ber o

f str

ains





4 Discussion









5 Conclusions




Acknowledgments


References
































































of




Disease Markers



OncologyJournal of





PPAR Research



Volume 2018


Journal of

ObesityJournal of




















Table1Con

tinued

Strain

no

Strain

name

GeneB

ankAc

cNo

Query

coverage

Maxident

Isolates

ourcea

80Weis

sella

confusaSYary1

KU324933

98

97

BMCM

81Weis

sella

confusaSho7ir

KU324934

96

98

BMCM

82Weis

sella

confusaFarag8

KU324935

97

97

BMCM

83Weis

sella

confusaA7G

afKU

324936

99

98

BMCM

92La

ctococcusgarvieaeZ

SJ5

KU324937

99

98

BMCM

93La

ctococcusgarvieaeE

mad4

KU324938

98

99

BMCM

a RCM

raw

cowmilk

RGMraw

goatmilk

RCA

Mraw

camelmilk

FCA

Mfrozencamelmilk

CMCM

cheesem

adefrom

cowmilk

CCM

CMcoo

kedcheese

madefrom

cowmilk

SCM

CMsalty

cheese

made

from

cowmilk

SYM

CMstirredyogu

rt(Laban)m

adefrom

cowmilk

SYM

CAMstirredyogu

rt(Laban)m

adefrom

camelmilk

YMCM

yogurtm

adefrom

cowmilk

BMCM

butterm

adefrom

cowmilk

QMCM

qeshta(cream

)madefrom

cowmilk

MMGMm

adheer

madefrom

goatmilk








Table2Th

eprofi

leof

theselected

LAB(n

=46

)for

mainprob

iotic

characteris

ticsa

ndtheantib

acteria

lactivity

spectraof

theirC

FSagainstseven

indicatoro

rganism

sAE

nterococcus

faecalisAT

CC29212B

EcoliAT

CC25922C

Salm

onellasppD

Shigella

sonn

eiAT

CC25931E

Staphylococcus

aureus

ATCC

25923F

MRS

AAT

CC43330

GL

isteria

monocytogenes

ATCC


n+

inhibitio

nzone

80ndash

100mm+

+inhibitio

nzone

101ndash

150mm+

++inh

ibition

zone

151ndash

200mm+

+++

inhibitio

nzone

200ndash300m

m

Strain

no-

code

Samples

ource

Strain

name

Accessionno

Hem

olysis

viabilityin

pH3medium

a

viabilityin

05

wv

bilea

Noof

antib

iotic

ssensitive

to15

AB

CD

EF

G

2RC

M2

Enterococcus

faecium

Rashad3

KU324893120574-hem

olysis

981plusmn0141

988plusmn0141

12+

++minus

+minus

++

3RC

M3

Enterococcus

durans

Etim

ad1

KU324894120574-hem

olysis

994plusmn0141

9775plusmn0354

12+

++minus

+minus

++

4RC

M4

Enterococcus

faecium

SMBM

3KU

324895120574-hem

olysis

991plusmn0283

981plusmn0283

13+

++minus

+minus

++

7CM

CM1

Lacto

bacilluscaseiMSJ1

KU324896120574-hem

olysis

9945plusmn0354

9905plusmn0212

9+

++++

+++

++

8CM

CM2

Lactobacillus

paracaseiN

MBM

1KU

324897120574-hem

olysis

9355plusmn0636

9845plusmn0212

10+

+++

+minus

+++

9CM

CM3

Lacto

bacilluscaseiBg

Shn3

KU324898120574-hem

olysis

9885plusmn0919

9925plusmn0212

9+

++

+minus

++

10CM

CM4

Lacto


7KU

324899120574-hem

olysis

975plusmn0283

974plusmn0424

9+

+++

+minus

+++

11CM

CM5

Lacto

bacilluscaseiDwan5

KU324900120574-hem

olysis

987plusmn0849

993plusmn0283

9+

+++

+++

++

12CM

CM6

Lactobacillus

plantarum

EyLan2

KU324901120574-hem

olysis

763plusmn0566

990plusmn0141

11+

++

++

++

19CC

MCM

1La

ctobacillus

futsa

iiEM

BM2

KU324902120574-hem

olysis

9835plusmn0778

918plusmn12

77

+++

+++minus

+++

23CC

MCM

5Weis

sella

confusaSaEd

-7KU

324903120572-hem

olysis

991plusmn0141

9545plusmn0354

10minus

minus+minusminus

+++

28RG

M5

Enterococcus

faecalisShbam40

KU324904120574-hem

olysis

822plusmn099

6745plusmn16

2612

minus++

++++

minusminus

+++

29RG

M6

Enterococcus

faecium

ZiNb3

KU324905120574-hem

olysis

994plusmn0141

996plusmn0141

12+

+minus

++

+++

32SY

MCM

3En

terococcus

faecium

Gail-B

awZir8

KU324906120574-hem

olysis

991plusmn0283

741plusmn2263

12++

+++

+++

++

34SY

MCM

5En

terococcus

faecium

NSJ2

KU324907120574-hem

olysis

9896plusmn0078

976plusmn0424

11+

++

++minus

+++

++35

SYMCM

6Streptococcustherm

ophilusB

inSlman8

KU324908120572-hem

olysis

6735plusmn14

859565plusmn0636

11minus

++minus

++++

minus+

36SY

MCM

7La

ctococcuslactis

HadRa

mi9

KU324909120574-hem

olysis

9895plusmn0212

655plusmn19

812

minus+

+++

minusminus

++37

QMCM

1En

terococcus

faecium

Marwh2

KU324910120574-hem

olysis

985plusmn0424

8365plusmn12

0212

++minus

+minus

++

38QMCM

2En

terococcus

faecium

Mon

a3KU

324911120574-hem

olysis

9755plusmn0354

6815plusmn16

2611

+++

++minus

+++

39QMCM

3En

terococcus

faecium

SSJ3

KU324912120574-hem

olysis

983plusmn0283

609plusmn12

7312

+++minus

+minus

++

40QMCM

4En

terococcus

lactisJedd

ah9

KU324913120574-hem

olysis

9715plusmn0354

6715plusmn0636

11minus

++

+minus

++

42QMC M

6Streptococcustherm

ophilusM

aNaL33

KU324914120572-hem

olysis

6085plusmn14

85951plusmn0424

11minus

++minus

++++

minus+

44MMGM1

Enterococcus

faecium

Adeb3

KU324915120574-hem

olysis

985plusmn0566

9735plusmn0636

8+

+++

++minus

minus++

45MMGM2

Enterococcus

faecium

ESJ4

KU324916120574-hem

olysis

991plusmn0283

598plusmn0283

12++

+minus

+minus

+++

46MMGM3

Enterococcus

durans

She7R

KU324917120574-hem

olysis

956plusmn0424

667plusmn12

7311

+++minus

+minus

+++

47MMGM4

Lactobacillus

acidophilusH

adhram

aut4

KU324918120574-hem

olysis

8265plusmn0636

907plusmn1131

10minus

+++

++minus

+++

50YM

CM3

Lactobacillus

acidophilusM

usallam2

KU324919120574-hem

olysis

9225plusmn0495

9945plusmn0212

11minus

+++

++minus

+++

+53

RCAM1

Enterococcus

faecium

BagH

om4

KU324920120574-hem

olysis

995plusmn01 41

9825plusmn0495

13+

++minus

+minus

++

58RC

AM6


Omer9

KU324921120574-hem

olysis

6455plusmn2758

975plusmn0283

13minus

++

++minus

minus++

60SY

MCA

M2


Anw

r4KU

324922120574-hem

olysis

7285plusmn12

029745plusmn0495

14minus

++

++minus

+++

++61

SYMCA

M3


Zaki1

KU324923120574-hem

olysis

7575plusmn19

09971plusmn0849

14minus

++

++minus

minus+

63SY

MCA

M5


Salam7

KU324924120574-hem

olysis

7445plusmn0636

9895plusmn0212

14minus

++

++minus

+++

+68

SCMCM

4En

terococcus

faecium

Ma7Fo

dKU

324925120574-hem

olysis

9845plusmn0636

377plusmn0990

13+

+++

++minus

++

73FC

AM4


JmaL3

KU324926120574-hem

olysis

5285plusmn0636

9555plusmn0636

13minus

+minusminus

++

++74

FCAM5


Foad7

KU324927120574-hem

o lysis

5675plusmn14

85953plusmn0566

12minus

+minusminus

++

++75

BMCM

1Weis

sella

confusaAhM

d8KU

324928120572-hem

olysis

9945plusmn0212

953plusmn0424

10minus

minus+minusminus

minusminus

76BM

CM2

Weis

sella

confusaTarim

4KU

324929120572-hem

olysis

9915plusmn0212

952plusmn0424

10minus

minus+minusminus

++++minus

77BM

CM3

Weis

sella

confusaNoo

R1KU

324930120572-hem

olysis

9955plusmn0071

9655plusmn0778

10++

++minusminus

+++minus

78BM

CM4

Weis

sella

confusaSaYu

n2KU

324931120572-hem

olysis

9945plusmn071

9635plusmn0495

10minus

++++minusminus

minusminus

79BM

CM5

Weis

sella

confusaMuK

alla5

KU324932120572-hem

olysis

709plusmn0707

9595plusmn0212

11minus

+minusminus

+++

+


Table2Con

tinued

Strain

no-

code

Samples

ource

Strain

name

Accessionno

Hem

olysis

viabilityin

pH3medium

a

viabilityin

05

wv

bilea

Noof

antib

iotic

ssensitive

to15

AB

CD

EF

G

80BM

CM6

Weis

sella

confusaSYary1

KU324933

120572-hem

olysis

992plusmn0141

982plusmn0424

11minus


+81

BMCM

7Weis

sella

confusaSho7ir

KU324934

120572-hem

olysis

969plusmn0566

949plusmn0849

10++

minus+

++

++

82BM

CM8

Weis

sella

confusaFarag8

KU324935

120572-hem

olysis

973plusmn0424

9565plusmn0636

10++


83BM

CM9

Weis

sella

confusaA7G

afKU

324936

120572-hem

olysis

9675plusmn0212

9515plusmn0636

10++

minus+minusminus

+minus

92BM

CM18


SJ5

KU324937

120572-hem

olysis

885plusmn0707

9865plusmn0354

11minus


93BM

CM19


mad4

KU324938

120572-hem

olysis

860plusmn0707

992plusmn0141

11minus


a Resultsarep

resented

asthem

eanvalueo

fdup

licatetria

lsplusmnstandard

deviation(SD)


3770 50-70 701-899 90-95 951-979 98-9955



0

5

10

15

20

25

Num

ber o

f str

ains





4 Discussion









5 Conclusions




Acknowledgments


References
































































of




Disease Markers



OncologyJournal of





PPAR Research



Volume 2018


Journal of

ObesityJournal of


























Table2Th

eprofi

leof

theselected

LAB(n

=46

)for

mainprob

iotic

characteris

ticsa

ndtheantib

acteria

lactivity

spectraof

theirC

FSagainstseven

indicatoro

rganism

sAE

nterococcus

faecalisAT

CC29212B

EcoliAT

CC25922C

Salm

onellasppD

Shigella

sonn

eiAT

CC25931E

Staphylococcus

aureus

ATCC

25923F

MRS

AAT

CC43330

GL

isteria

monocytogenes

ATCC


n+

inhibitio

nzone

80ndash

100mm+

+inhibitio

nzone

101ndash

150mm+

++inh

ibition

zone

151ndash

200mm+

+++

inhibitio

nzone

200ndash300m

m

Strain

no-

code

Samples

ource

Strain

name

Accessionno

Hem

olysis

viabilityin

pH3medium

a

viabilityin

05

wv

bilea

Noof

antib

iotic

ssensitive

to15

AB

CD

EF

G

2RC

M2

Enterococcus

faecium

Rashad3

KU324893120574-hem

olysis

981plusmn0141

988plusmn0141

12+

++minus

+minus

++

3RC

M3

Enterococcus

durans

Etim

ad1

KU324894120574-hem

olysis

994plusmn0141

9775plusmn0354

12+

++minus

+minus

++

4RC

M4

Enterococcus

faecium

SMBM

3KU

324895120574-hem

olysis

991plusmn0283

981plusmn0283

13+

++minus

+minus

++

7CM

CM1

Lacto

bacilluscaseiMSJ1

KU324896120574-hem

olysis

9945plusmn0354

9905plusmn0212

9+

++++

+++

++

8CM

CM2

Lactobacillus

paracaseiN

MBM

1KU

324897120574-hem

olysis

9355plusmn0636

9845plusmn0212

10+

+++

+minus

+++

9CM

CM3

Lacto

bacilluscaseiBg

Shn3

KU324898120574-hem

olysis

9885plusmn0919

9925plusmn0212

9+

++

+minus

++

10CM

CM4

Lacto


7KU

324899120574-hem

olysis

975plusmn0283

974plusmn0424

9+

+++

+minus

+++

11CM

CM5

Lacto

bacilluscaseiDwan5

KU324900120574-hem

olysis

987plusmn0849

993plusmn0283

9+

+++

+++

++

12CM

CM6

Lactobacillus

plantarum

EyLan2

KU324901120574-hem

olysis

763plusmn0566

990plusmn0141

11+

++

++

++

19CC

MCM

1La

ctobacillus

futsa

iiEM

BM2

KU324902120574-hem

olysis

9835plusmn0778

918plusmn12

77

+++

+++minus

+++

23CC

MCM

5Weis

sella

confusaSaEd

-7KU

324903120572-hem

olysis

991plusmn0141

9545plusmn0354

10minus

minus+minusminus

+++

28RG

M5

Enterococcus

faecalisShbam40

KU324904120574-hem

olysis

822plusmn099

6745plusmn16

2612

minus++

++++

minusminus

+++

29RG

M6

Enterococcus

faecium

ZiNb3

KU324905120574-hem

olysis

994plusmn0141

996plusmn0141

12+

+minus

++

+++

32SY

MCM

3En

terococcus

faecium

Gail-B

awZir8

KU324906120574-hem

olysis

991plusmn0283

741plusmn2263

12++

+++

+++

++

34SY

MCM

5En

terococcus

faecium

NSJ2

KU324907120574-hem

olysis

9896plusmn0078

976plusmn0424

11+

++

++minus

+++

++35

SYMCM

6Streptococcustherm

ophilusB

inSlman8

KU324908120572-hem

olysis

6735plusmn14

859565plusmn0636

11minus

++minus

++++

minus+

36SY

MCM

7La

ctococcuslactis

HadRa

mi9

KU324909120574-hem

olysis

9895plusmn0212

655plusmn19

812

minus+

+++

minusminus

++37

QMCM

1En

terococcus

faecium

Marwh2

KU324910120574-hem

olysis

985plusmn0424

8365plusmn12

0212

++minus

+minus

++

38QMCM

2En

terococcus

faecium

Mon

a3KU

324911120574-hem

olysis

9755plusmn0354

6815plusmn16

2611

+++

++minus

+++

39QMCM

3En

terococcus

faecium

SSJ3

KU324912120574-hem

olysis

983plusmn0283

609plusmn12

7312

+++minus

+minus

++

40QMCM

4En

terococcus

lactisJedd

ah9

KU324913120574-hem

olysis

9715plusmn0354

6715plusmn0636

11minus

++

+minus

++

42QMC M

6Streptococcustherm

ophilusM

aNaL33

KU324914120572-hem

olysis

6085plusmn14

85951plusmn0424

11minus

++minus

++++

minus+

44MMGM1

Enterococcus

faecium

Adeb3

KU324915120574-hem

olysis

985plusmn0566

9735plusmn0636

8+

+++

++minus

minus++

45MMGM2

Enterococcus

faecium

ESJ4

KU324916120574-hem

olysis

991plusmn0283

598plusmn0283

12++

+minus

+minus

+++

46MMGM3

Enterococcus

durans

She7R

KU324917120574-hem

olysis

956plusmn0424

667plusmn12

7311

+++minus

+minus

+++

47MMGM4

Lactobacillus

acidophilusH

adhram

aut4

KU324918120574-hem

olysis

8265plusmn0636

907plusmn1131

10minus

+++

++minus

+++

50YM

CM3

Lactobacillus

acidophilusM

usallam2

KU324919120574-hem

olysis

9225plusmn0495

9945plusmn0212

11minus

+++

++minus

+++

+53

RCAM1

Enterococcus

faecium

BagH

om4

KU324920120574-hem

olysis

995plusmn01 41

9825plusmn0495

13+

++minus

+minus

++

58RC

AM6


Omer9

KU324921120574-hem

olysis

6455plusmn2758

975plusmn0283

13minus

++

++minus

minus++

60SY

MCA

M2


Anw

r4KU

324922120574-hem

olysis

7285plusmn12

029745plusmn0495

14minus

++

++minus

+++

++61

SYMCA

M3


Zaki1

KU324923120574-hem

olysis

7575plusmn19

09971plusmn0849

14minus

++

++minus

minus+

63SY

MCA

M5


Salam7

KU324924120574-hem

olysis

7445plusmn0636

9895plusmn0212

14minus

++

++minus

+++

+68

SCMCM

4En

terococcus

faecium

Ma7Fo

dKU

324925120574-hem

olysis

9845plusmn0636

377plusmn0990

13+

+++

++minus

++

73FC

AM4


JmaL3

KU324926120574-hem

olysis

5285plusmn0636

9555plusmn0636

13minus

+minusminus

++

++74

FCAM5


Foad7

KU324927120574-hem

o lysis

5675plusmn14

85953plusmn0566

12minus

+minusminus

++

++75

BMCM

1Weis

sella

confusaAhM

d8KU

324928120572-hem

olysis

9945plusmn0212

953plusmn0424

10minus

minus+minusminus

minusminus

76BM

CM2

Weis

sella

confusaTarim

4KU

324929120572-hem

olysis

9915plusmn0212

952plusmn0424

10minus

minus+minusminus

++++minus

77BM

CM3

Weis

sella

confusaNoo

R1KU

324930120572-hem

olysis

9955plusmn0071

9655plusmn0778

10++

++minusminus

+++minus

78BM

CM4

Weis

sella

confusaSaYu

n2KU

324931120572-hem

olysis

9945plusmn071

9635plusmn0495

10minus

++++minusminus

minusminus

79BM

CM5

Weis

sella

confusaMuK

alla5

KU324932120572-hem

olysis

709plusmn0707

9595plusmn0212

11minus

+minusminus

+++

+


Table2Con

tinued

Strain

no-

code

Samples

ource

Strain

name

Accessionno

Hem

olysis

viabilityin

pH3medium

a

viabilityin

05

wv

bilea

Noof

antib

iotic

ssensitive

to15

AB

CD

EF

G

80BM

CM6

Weis

sella

confusaSYary1

KU324933

120572-hem

olysis

992plusmn0141

982plusmn0424

11minus


+81

BMCM

7Weis

sella

confusaSho7ir

KU324934

120572-hem

olysis

969plusmn0566

949plusmn0849

10++

minus+

++

++

82BM

CM8

Weis

sella

confusaFarag8

KU324935

120572-hem

olysis

973plusmn0424

9565plusmn0636

10++


83BM

CM9

Weis

sella

confusaA7G

afKU

324936

120572-hem

olysis

9675plusmn0212

9515plusmn0636

10++

minus+minusminus

+minus

92BM

CM18


SJ5

KU324937

120572-hem

olysis

885plusmn0707

9865plusmn0354

11minus


93BM

CM19


mad4

KU324938

120572-hem

olysis

860plusmn0707

992plusmn0141

11minus


a Resultsarep

resented

asthem

eanvalueo

fdup

licatetria

lsplusmnstandard

deviation(SD)


3770 50-70 701-899 90-95 951-979 98-9955



0

5

10

15

20

25

Num

ber o

f str

ains





4 Discussion









5 Conclusions




Acknowledgments


References
































































of




Disease Markers



OncologyJournal of





PPAR Research



Volume 2018


Journal of

ObesityJournal of




















Table2Con

tinued

Strain

no-

code

Samples

ource

Strain

name

Accessionno

Hem

olysis

viabilityin

pH3medium

a

viabilityin

05

wv

bilea

Noof

antib

iotic

ssensitive

to15

AB

CD

EF

G

80BM

CM6

Weis

sella

confusaSYary1

KU324933

120572-hem

olysis

992plusmn0141

982plusmn0424

11minus


+81

BMCM

7Weis

sella

confusaSho7ir

KU324934

120572-hem

olysis

969plusmn0566

949plusmn0849

10++

minus+

++

++

82BM

CM8

Weis

sella

confusaFarag8

KU324935

120572-hem

olysis

973plusmn0424

9565plusmn0636

10++


83BM

CM9

Weis

sella

confusaA7G

afKU

324936

120572-hem

olysis

9675plusmn0212

9515plusmn0636

10++

minus+minusminus

+minus

92BM

CM18


SJ5

KU324937

120572-hem

olysis

885plusmn0707

9865plusmn0354

11minus


93BM

CM19


mad4

KU324938

120572-hem

olysis

860plusmn0707

992plusmn0141

11minus


a Resultsarep

resented

asthem

eanvalueo

fdup

licatetria

lsplusmnstandard

deviation(SD)


3770 50-70 701-899 90-95 951-979 98-9955



0

5

10

15

20

25

Num

ber o

f str

ains





4 Discussion









5 Conclusions




Acknowledgments


References
































































of




Disease Markers



OncologyJournal of





PPAR Research



Volume 2018


Journal of

ObesityJournal of




















3770 50-70 701-899 90-95 951-979 98-9955



0

5

10

15

20

25

Num

ber o

f str

ains





4 Discussion









5 Conclusions




Acknowledgments


References
































































of




Disease Markers



OncologyJournal of





PPAR Research



Volume 2018


Journal of

ObesityJournal of

























5 Conclusions




Acknowledgments


References
































































of




Disease Markers



OncologyJournal of





PPAR Research



Volume 2018


Journal of

ObesityJournal of












































































of




Disease Markers



OncologyJournal of





PPAR Research



Volume 2018


Journal of

ObesityJournal of



















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