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Activ i Dad Cane Lapol Voc on Cent

Activ i Dad Cane Lapol Voc on Cent

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Actividad antimicrobiana de la canela en polvo
Actividad antimicrobiana de la canela en polvo

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Published by: chamacos on Sep 15, 2012
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GRANULARITY AND ANTIBACTERIAL ACTIVITIES OF ULTRA-FINECINNAMON AND CLOVE POWDERS
 jfs_300 291..296
XUANKUANG,BINLI,
1
RUIKUANG,XIAODONGZHENG,BOZHU,BAOLIXU,MEIHUMA
College of Food Science and Technology, Huazhong Agricultural University, Wuhan 430070, China
1
Corresponding author. TEL:
+
86-1329-6597469; FAX:
+
86-27-87282966;EMAIL: libin343@yahoo.com.cnAccepted for Publication December 9, 2010doi:10.1111/j.1745-4565.2011.00300.x
ABSTRACT
This article describes a novel and simple method that using ultra-fine powders of ball-milledcinnamonandclovedirectlytoinhibitthegrowthof fivespoilageorgan-isms from meat. The effect of ball-milled time on powder granularity and the par-ticle sizes on the antimicrobial effectiveness were evaluated. The results from theantibacterial tests demonstrated that the ultra-fine powders had strong antimicro-bial activities, the minimum inhibitory concentrations (MICs) of the clove powderand the cinnamon powder were from 1.0 to 1.5% (w/v) against
Escherichia coli
,
Sta- phylococcus aureus
,
Brochothrix thermosphacta and Lactobacillus rhamnosus
,
and 
thesame against
Pseudomonas fluorescens
were 2.0 and 2.5%, respectively. The inhibi-toryeffectincreasedwithincreasingpowderconcentrationsfrom0.5to2.5%(w/v).However, obvious reversing characteristic appeared when the particle sizesdecreased to 7.085
m
m for cinnamon and 8.915
m
m for clove. This study suggestedthat traditional spice powders with proper particle sizes taking the place of essentialoils as effective antibacterials had a wide application prospect for their low cost andsafety.
PRACTICAL APPLICATIONS
Ball mills have been successfully used for grinding raw materials to ultra-finepowders because of their easier operation, higher grinding rate, lower energy con-sumptionandconsiderablylessplantspace.However,beforecarryingoutthisstudy,it was unknown whether ultra-fine powders of ball-milled cinnamon and clove candirectlyinhibitthegrowthoffivespoilageorganismsfrommeat.Inthepresentwork,the effect of ball-milled time on the powder granularity and the particle sizes on theantimicrobial effectiveness were studied. This study showed that both traditionalspice powders with proper particle sizes taking the place of essential oils as effectiveantibacterials had strong antimicrobial activities.
INTRODUCTION
The spoilage of raw and processed food is generally attrib-uted to the action of bacteria (Borch
et al 
. 1996). At present,a number of studies have demonstrated that both the cinna-mon (
Cinnamomum cassia
) oil and clove (
Eugenia caryo- phyllus
) oil had a broad range of antimicrobial activitiesagainst gram-positive and gram-negative bacteria (Moreira
et al 
. 2005; López-Malo
et al 
. 2007; Gupta
et al 
. 2008), and ahigh fungicidal affects against molds, yeasts (Conner andBeuchat 1984) and postharvest decay fungi of grape:
 Aspergillus niger 
,
Alternaria alternata
,
Colletotrichum gloeosporioides
,
Lasiodiplodia theobromae
,
Phomopsis viticola
and
Rhizopus stolonifer 
(Sukatta
et al 
. 2008).Cinnamon and clove are traditional food spices and arecommonly used in the food industry because of their specialaroma as well as their antibacterial and antifungal activities(Bakkali
et al 
. 2008; Kechichian
et al 
. 2010). The aroma andantibacterial activities are most attributed to a number of small terpenoid and phenolic compounds in them (Beuchat2000; Vattem
et al 
. 2004). Generally speaking, the activeingredientsof herbsmainlylocatedinthecells.Incaseswhereplantsareusedintheirnaturalstate,theeffectivecomponentsare used only after crossing cell’s outer membranes.If the cell
Journal of Food Safety ISSN 1745-4565
291
Journal of Food Safety
31
(2011) 291–296 © 2011 Wiley Periodicals, Inc.
 
walls were broken, the active ingredients would contact thefoodandthereforebecomemorebio-available.However,tra-ditional solvent extraction of essential oil requires a largequantityof toxicorganicsolvent.Atthesametime,itislabor-intensive, requires long extraction time, and has low extrac-tion yield. Thus the direct use of ultra-fine powders of foodspices to replace essential oil should be explored.Ball mills have been successfully used for grinding raw materials into ultra-fine powders because of their ease of operation, higher grinding rate, lower energy consumptionand considerably less plant space compared with other fine-grinding machines (Jimbo 1992; Gao and Forssberg 1995).This can reduce the particle sizes of plants down to 10
m
m orlower (Lowrison 1974). However, to the best of our knowl-edgethereisnoliteratureinvestigatingthefeasibilityof ultra-fine powders of spices as potential antimicrobial agents. Inthis work, we grinded cinnamon and clove using a high-frequency oscillatory type ball mill, and examined the anti-microbialactivitiesobtainedtheultra-finepowdersof cinna-mon and clove against five spoilage organisms
E. coli
,
S.aureus
,
B.thermosphacta
,
L.rhamnosusandP.fluorescens
frommeats.Theeffectof ball-milledtimeonthepowdergranular-ity and the particle sizes on the antimicrobial effectivenesswere evaluated.
MATERIALS AND METHODS
Preparation of Bacterial Strains
Escherichia coli
(strain AS1.1366),
Staphylococcus aureus
(strain GIM1.55),
Pseudomonas fluorescens
(strain AS1.823)were obtained from Microbial Culture Collection Center of Guangdong Institute of Microbiology (Guangdong, China).
Brochothrix thermosphacta
(1A01916) was provided by MarineCultureCollectionofChina(Fujian,China).
Lactoba-cillus rhamnosus
(CICC 6141) was obtained from ChinaCenter of Industrial Culture Collection (Beijing, China).Man–Rogosa–Sharpe (MRS) medium and streptomycinthalliumacetate actidion (STA) medium were purchasedfrom Qingdao Hope Bio-Technology Co., Ltd. (Qingdao,China).
E. coli
,
S. aureus
and
P. fluorescens
were suspended in asterile nutrient broth (NB) medium, and incubated at37
Ϯ
1C, 37
Ϯ
1C and 30
Ϯ
1C for 24 h, respectively.
L.rhamnosus
was transferred into a sterile MRS medium andincubated at 37
Ϯ
1C for 48 h,whereas
B. thermosphacta
wastransferred to a sterile STA medium and incubated at30
Ϯ
1C for 48 h. One milliliter stock culture was standard-ized through two successive 24 h growth cycles in an appro-priate broth (NB, MRS or STA). The cell from thestandardized culture was inoculated in a sterile medium andincubated to obtain a cell suspension at exponential phasecontaining approximately 2
¥
10
7
cfu/mL.
Preparations and Characterizations ofUltra-Fine Powders
Cinnamon (
Cinnamomum cassia
) and clove (
Eugenia caryo- phyllus
) were purchased from Jinyaotang Drugstore inWuhan, Hubei Province, China. The samples were dried at50C for 24 h, and then milled for 50, 60, 70, 80 and 90 minusing a high-frequency oscillatory type ball mill at 20
Ϯ
1C.The cinnamon powders were coded as CI-50, CI-60, CI-70,CI-80 and CI-90, the clove powders as CL-50, CL-60, CL-70,CL-80 and CL-90,respectively.The particle size of a sample was analyzed using a particlesize analyzer (LS 13320, Deckman Coulter Inc., Fullerton,CA)andthemeanparticlesize(d
50
)ofasamplewasestimatedas a median diameter from the count value based on weightpercent cumulative curve.
Determination of Antibacterial Activities
The antibacterial assays of cinnamon and clove powdersagainst selected
E. coli
,
S. aureus
,
P. fluorescens
,
L. rhamnosus
and
B. thermosphacta
were determined by broth dilutionmethod.Eachtestedbacteriumwasculturedinacorrespond-ing medium described above. A series of broths containingtested powders were prepared, the concentrations of eachtested powder incorporated in broth were 0,50,100,150,200and 250 mg/mL. Each bacterium was suspended in sterilewater and diluted to approximately 2
¥
10
7
cfu/mL. Then1 mL bacterial suspension was added to 9 mL broth contain-ing a sample, and the mixture was cultured at 37C for 48 hwith stirring to prevent the sample from depositing.The sur-vivingbacteriawereconfirmedbyplatecount.Theinhibitory effect (I %) was calculated using the following formula:
A B A
% %
=×
100(1)where
 A
isthenumbersof bacteriawithoutanysampleinthetest tube;
B
is the numbers of bacteria with sample in the testtube.Three independent measurements were prepared in eachcase. The MIC of a sample was defined as the lowest concen-tration of the sample in which more than 90% bacteria couldnot grow.
Statistical Analysis
Data were subjected to analysis of variance and Duncan’smultiple-range tests using analysis of variance procedure.Differencesbetweenmeansof bacterialcounts(cfu/mL)wereregarded to be statistically significant when
<
0.05. Experi-ments were replicated twice for each powder sample at eachconcentration.
GRANULARITYANDANTIBACTERIALACTIVITIES
X. KUANG
ET AL
.
292
Journal of Food Safety
31
(2011) 291–296 © 2011 Wiley Periodicals, Inc.
 
RESULTS AND DISCUSSION
Granularity
The particle size distributions of cinnamon and clovepowderswithdifferentball-milledtimewereshowninFig. 1.The diameter of ball-milled clove powder for 50 min was inthe wide range of 5.01–338.84
m
m. When ball-milled timeincreased, the range narrowed down, while when the ball-milled time was 90 min, the distribution was from 5.25 to30.31
m
m. The volume distribution of clove powder ball-milled for 50 min showed the typical four populations withpeak values in 10.10, 27.54, 100.89 and 213.80
m
m, respec-tively.However,it gradually turned to three,two populationsand even almost a single peak with increasing ball-milledtime, which means the particle size become more symmetri-cal. As for cinnamon, the distribution range of particle sizealso narrowed down when the ball-milled time was increas-ing, but it exhibited a single peak at all the different milledtimes.The d
50
values of all the samples with different ball-milledtimewereshowninFig. 2.Itindicatedthattheparticlesizesof both cinnamon powder and clove powder decreased rapidly duringball-milled50–60 min,howeveritdecreasedrelatively slow in the next 10 min. Seen also in Fig. 1, the particle sizedistributions of CI60 and CI70 (CL60 and CL70) were very similar. It suggested that this period might be the stalematestageforcelldisruption.Afterthis,theparticlesizesdecreasedsharplyagaininthefollowing10 min,thed
50
valuesof cinna-mon and clove powders decreased to 11.19 and 14.39
m
m,respectively. In the last 10 min, the particle size decreasingratesbecamesmalleronceagain,untilfinallyalmostceasedtochange. The d
50
values of CI90 and CL90 were 7.085 and8.815
m
m,respectively.
MICs
The inhibitory effect of cinnamon and clove powders againstfive bacterial strains with concentrations ranging from 0.5 to2.5%waspresentedinFig. 3.Thechosencinnamonandclovepowders were CI-70 (d
50
=
17
m
m) and CL-70 (d
50
=
21
m
m),which were at intermediate levels of five different particlesizes. The results showed that both kind of powers signifi-cantlyreducedthetestedmicroorganismlevelsattheconcen-tration
Յ
2.5% (w/v) (
<
0.05). The inhibitory effect
CI-50CI-60CI-70CI-80 CI-90CL-50CL-60CL-70CL-80CL-90
FIG. 1.
PARTICLE SIZE DISTRIBUTIONS OFCINNAMON AND CLOVE POWDERSBALL-MILLED WITH DIFFERENT TIMECI, cinnamon; CL, clove; 50, 60, 70, 80and 90, milled for 60, 70, 80 and 90 min,respectively.
01428425670CinnmonClove
   M  e  a  n  p  a  r   t   i  c   l  e  s   i  z  e  s   (  u  m   )
50 60 70 80 90Ball-milled time (min)
FIG. 2.
EFFECT OF BALL-MILLED TIME ON MEAN PARTICLE SIZE (d
50
) OFCINNAMON AND CLOVE POWDERSX. KUANG
ET AL
.
GRANULARITYANDANTIBACTERIALACTIVITIES
293
Journal of Food Safety
31
(2011) 291–296 © 2011 Wiley Periodicals, Inc.

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