حلمي القدسيمشرف قسم صناعات الاغذية
عدد المساهمات : 350
نقاط : 5300
تاريخ التسجيل : 03/09/2011
العمر : 35
 | موضوع: Blood Agar Base No. 2 M834  الثلاثاء يوليو 10, 2012 11:43 am | |
| Blood Agar Base No. 2 M834Blood Agar Base No. 2 is specially
devised to permit the maximum recovery of streptococci, pneumococci and otherfastidious pathogenic microorganisms
without interfering with their haemolytic reactions. Composition**Ingredients Gms / LitreProteose peptone 15.000Liver extract 2.500Yeast extract 5.000Sodium chloride 5.000Agar 15.000Final pH ( at 25°C) 7.4±0.2**Formula adjusted, standardized to suit
performance parameters DirectionsSuspend 21.25 grams in 500 ml distilled
water. Heat to boiling to dissolve the medium completely. Sterilize by
autoclaving at15 lbs pressure (121°C) for 15 minutes.
Cool to 40 - 50°C and aseptically add 7% v/vsterile defibrinated blood.For Brucella species: Add
rehydrated contents of 1 vial of Brucella Selective Supplement (FD005) to500 ml sterile molten base.For Campylobacter species : Add
rehydrated contents of 1 vial of Campylobacter Supplement - I (FD006) or
CampylobacterSupplement - II (FD007) or Campylobacter
Supplement - III (FD008) or Campylobacter Growth Supplement (FD009) to 500ml sterile molten base.For Streptococcus species: Add
rehydrated contents of 1 vial of Strepto Supplement (FD031) to 500 ml sterile
molten base.Mix well and pour into sterile Petri
plates. Principle And InterpretationA fastidious organism is one with
complete nutritional requirements, needing additional cellular building-block
molecules inorder to survive (1). Blood Agar Base
No. 2 is a highly nutritive medium. Microorganisms producing haemolysin give
visiblehaemolytic zones on this medium. It also
serves as a differential medium for Brucella and Campylobacter species
byadding different antibiotic supplements
for the respective bacteria (2, 3). Brucella cultures are highly
infective and must behandled with care. Incubate preferably
in 5-10% carbon dioxide atmosphere. Comparative studies of horse, rabbit and
sheepblood showed that sheep blood gave the
clearest and most reliable colony and haemolysis characteristics at both 24 and
48hours of incubation (4). It can be used
to prepare Chocolate Agar for the isolation of Haemophilus and Neisseria
species.It can also be used for primary
isolation of Haemophilus species, where horse blood is used for
enrichment. Better results areobtained by spreading half of the horse
blood agar plate with 2 drops of 10% saponin (5). Liver extract and yeast
extract helpsenhance the growth and haemolytic
reactions of fastidious organisms like Streptococci and Pneumococci. Proteose
peptoneserves as the nitrogen source while
liver digest and yeast extract provide essential carbon, vitamin, nitrogen and
amino acidsources. Sodium chloride maintains the
osmotic equilibrium. Supplementation with blood (5-10%) provides additional
growthfactors and also serves as basis for
determining haemolytic reactions. Haemolytic patterns may vary with the source
of animalblood or type of base medium used (6). Quality ControlAppearanceCream to yellow homogeneous free flowing
powderGellingFirm, comparable with 1.5% Agar gel HiMedia Laboratories Technical DataDisclaimer :User must ensure suitability of the
product(s) in their application prior to use. Products conform solely to the
information contained in thisand other related HiMedia™ publications. The
information contained in this publication is based on our research and
development workand is to the best of our knowledge true and
accurate. HiMedia™ Laboratories Pvt Ltd reserves the right to make changes to
specificationsand information related to the products at
any time. Products are not intended for human or animal diagnostic or
therapeutic use but forlaboratory, research or further manufacturing
use only, unless otherwise specified. Statements contained herein should not be
consideredas a warranty of any kind, expressed or
implied, and no liability is accepted for infringement of any patents.HiMedia Laboratories Pvt. Ltd. A-516,Swastik
Disha Business Park,Via Vadhani Ind. Est., LBS Marg, Mumbai-400086, India.
Customer care No.: 022-61471919 Email: techhelp@himedialabs.comColour and Clarity of prepared mediumBasal medium : Yellow coloured clear to
slightly opalescent gel. After addition of 5-7% v/v sterile defibrinated blood:Cherry red coloured opaque gel forms in
Petri plates.ReactionReaction of 4.25% w/v aqueous solution
at 25°C. pH : 7.4±0.2pH7.20-7.60Cultural ResponseCultural characteristics observed with
added 5-7% sterile defibrinated blood, after an incubation at 35-37°C for 18-48
hours.Cultural ResponseOrganism Inoculum(CFU)Growth Recovery HaemolysisCultural ResponseNeisseria meningitidis ATCC1309050-100 good-luxuriant >=70% noneStaphylococcus aureusATCC 2592350-100 good-luxuriant >=70% betaStreptococcus pneumoniaeATCC 630350-100 good-luxuriant >=70% alphaStreptococcus pyogenesATCC 1961550-100 good-luxuriant >=70% betaStorage and Shelf LifeStore below 30°C in tightly closed
container and the prepared medium at 2-8°C. Use before expiry date on the
label.Reference1. Norton C. F., 1986, Microbiology, 2nd
Edition, Addison-Wesley Publishing Company.2. Hunter D. and Kearns M., 1977, Brit.
Vet. J., 133:486.3. Skirrow M. B., 1977, B.M.J., ii: 9.4. Snavely and Brahier, 1960, Am. J.
Clin. Pathol., 33:511.5. Waterworth and Pamela M., 1955, Brit.
J. Exp. Pathol., 36:186.6. Murray P. R,, Baron E. J., Jorgensen
J. H., Pfaller M. A., Yolken R. H., (Eds.), 8th Ed., 2003, Manual of ClinicalMicrobiology, ASM, Washington, D.C.Revision : 1 / 2011 |
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