DOCSLIB.ORG

Hemolytic Properties of the Mastitis Streptococcus* P

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Hemolytic Properties of the Mastitis Streptococcus* P Hemolytic Properties of the Mastitis Streptococcus* P. ARNE HANSEN, G. J. HUCKER, PH.D., AND MARION A. SNYDER New York State Agricultural Experiment Station, Geneva, N. Y. BLOOD agar plates have gained wide tion, be seen to contain the red blood use in the identification of bacteria, corpuscles in a fixed condition. The especially in the case of streptococci. surface colony should not be used in the Introduced by Marmorek (1902) and classification as it may be indistinguish- later, studied more in detail by Schott- able from the beta type. muller (1903), the use of blood agar The beta type is characterized by the has come to be a useful procedure in surrounding halo which is perfectly identifying streptococci. Brown (1919) clear. No red blood corpuscles may be described in detail the various types of seen in this clear zone even when ex- colonies produced on solid blood agar amined microscopically by magnifica- and emphasized the importance of using tion. The gamma type is produced by standard procedures and a standard organisms which do not produce means of designation. He divided the hemolysis or decolorize the blood various types of colonies into groups, medium. Brown calls both the alpha viz., alpha, alpha prime, beta and prime and the beta type, " hemolytic," gamma, these terms to apply only to yet the former is considered more closely deep colonies. The alpha type produces related to the alpha than to the beta green coloration of the medium sur- type. rounding the colonies. Under low The blood agar plate has been used power magnification faint hemolysis widely in the case of the mastitis strep- may be noted in the outer portions of tococcus. By a study of the literature the zone. When a plate seeded with it may be noted that no agreement is alpha hemolytic colonies is placed in the found regarding the hemolytic powers refrigerator after previous incubation of of this streptococcus. The typical 48 hours at 370 C., an outer hemolytic streptococcus of mastitis (Streptococcus zone will be noticed. The alpha prime mastitidis) t is as a rule described as type colony is intermediate between the being alpha or gamma hemolytic, but alpha and the beta types. The colony not infrequently reports are found is surrounded by a hemolyzed zone. designating it as beta hemolytic. This zone, however, is not clear and may, Gminder (1912) tested 26 udder when viewed under low power magnifica- streptococci on the blood plate prepared t In this discussion the term Streptococcus mastitidis * Read before the Laboratory Section of the Migula will be used to designate the streptococcus as- American Public Health Association at the Sixty-second sociated with chronic or subclinical mastitis. The Annual Meeting in Indianapolis, Ind., October 12, nomenclature of this organism will be subsequently 1933. determined. The use of this terminology at this time Approved by the Director of the New York State should not be construed to infer that the authors Agricultural Experiment Station as Journal Paper prefer this name to Streptococcus agalactiae Lehmann No. 8. and Neumann. t1262] MASTITIS STREPTOCOCCUS 1263 according to Schottmiiller. Four were ference in composition of the blood hemolytic. medium used. Haupt added 0.05 per Puppel (1912) found that out of 18 cent glucose while they used a sugar- mastitis streptococci, only 1 was hemo- free medium. lytic on human blood agar. Diernhofer (1932) mentions two Jones (1918) described hemolysis in varieties of S. mastitidis, one as an- a number of strains which very prob- hemolytic and the other as a beta ably could be termed S. mastitidis, but hemolytic variety. he states: " Mastitis caused by infec- Seelemann (1932) did not observe tions with non-hemolytic streptococci is hemolysis in true mastitis streptococci. more prevalent than that caused by He tested 300 strains using red cells of other classes of micro6rganisms." ox and sheep and 80 on horse blood Ayers and Mudge (1922) examined agar. In some cases a faint clear halo streptococci originating from the udder. was observed. Steck (1932) examined Of the 79 strains which were found to 24 S. mastitidis strains on Brown plates be S. mastitidis, 64 were of the beta and found that all were of the alpha hemolytic type, 15 of the gamma type. type. Hadley and Frost (1933) use The blood medium used was prepared the name S. mitis for the alpha strepto- according to Brown and no sugar was coccus responsible for the majority of added. cases of mastitis. They reserve the Carpenter (1925) found in 150 term S. mastitidis for beta hemolytic samples of milk from suspected cases of types. mastitis, alpha hemolytic streptococci in Rosell (1933) examined the be- 79 per cent and beta hemolytic in 6.6 havior of 268 strains of typical mastitis per cent of the cases. Whether the streptococci and found 3 only to be beta latter were S. mastitidis in a strict sense hemolytic, the remaining being of the is impossible to say from the data alpha or gamma types, using horse and presiented. sheep blood in the Brown technic. On Minett, Stableforth and Edwards cow's blood 10 per cent of the strains (1929) found more than half of the were beta hemolytic. No sugar was strains they considered to be S. masti- added to the medium, but horse meat or tidis bovis to be beta hemolytic and Liebig's meat extract were recom- the majority of the remainder. to be of mended. the viridans type. The disagreement between different Bergey (1930) gives S. mastitidis as authors in regard to the action of S. hemolytic. mastitidis toward blood media may be Klimmer and Haupt state (1930): due to several reasons: (1) The name "S. agalactiae grows, in our experience S. mastitidis, may be taken in too predominantly as the alpha type, rarely broad a sense including practically any according to the alpha prime type, com- streptococcus that may be isolated paratively frequently as the gamma from the udder. (2) Differences in type, never as the beta type." technic, composition of the medium, Haupt (1931) never found beta kind of blood, observation of surface hemolysis in S. mastitidis. colonies being the most important. Minett, Stableforth and Edwards (3) Difference in interpretation. Alpha (1931) stated in answer to Haupt that prime colonies are, no doubt, frequently S. mnastitidis may be either alpha or regarded as beta colonies. beta hemolytic and pointed out that The object of this discussion is to the variation in results between Haupt present data regarding the action on and themselves was caused by a dif- blood of authentic strains of S. masti- 1264 AMERICAN JOURNAL OF PUBLIC HEALTH tidis and the possible significance of Distinct and vigorous beta hemolysis this characteristic in the detection of was never observed, regardless of the udder infections. kind of blood or the presence or ab- sence of glucose. This observation is TECHNIC especially significant when it is realized Freshly grown cultures were plated in that some workers report that the same the various media and examined after strain varies in different laboratories 48 hours' incubation at 3 70 C. The from alpha to strong beta hemolytic. plates were then placed in the refrigera- The types beta, alpha, alpha prime, tor, chilled for another 48 hours, and and gamma may not represent distinct examined again. The media used were: types in all cases and further study is (1) horse blood agar, prepared from fer- needed to decide on the mechanism of mented veal broth, (2) horse blood agar, hemolysis in the weak beta hemolytic prepared from plain veal broth, (3) strains. Hemolysis is a complicated horse blood agar, prepared from plain process depending on many factors of veal broth plus 0.05 per cent glucose, which not all are fully understood. The (4) sheep blood agar, prepared from quantitative importance of these various plain veal broth plus 0.05 per cent factors has never been worked out; glucose, (5) cow blood agar prepared productions of true hemolysins, of acid from plain veal broth. The broth which and of peroxide. It is probable that constituted the basic part of all the the various types of hemolysis for the media had the following composition: time being should only be considered Veal infusion from 500 g. veal.... 1,000 c.c. as rough descriptions which should be Bacto peptone ................... 10 g. confirmed by quantitative measure- NaCl ....................... 5 g. ments. Agar . ...................... 15 g. While it may be difficult or impos- Reaction adjusted to pH 7.4 sible to distinguish between the various The media were tubed in 12 c.c. lots types, the difficulty, frequently- de- and 0.6 c.c. of blood was added before scribed, that a certain strain varies use. The blood was always left at least greatly from time to time when tested 48 hours in the cold before being used on blood plates has not been en- for plating. countered. Different transfers of the To supplement the results as secured same strain kept for years in different with the blood agar plates, tests for culture collections under varied con- hemolysin production were carried out ditions gave identical results. Many in liquid media. One-half c.c. of an 18 examples of this type are found in these hour culture in sugar-free broth was data (see 1 and 17, 3 and 8, 6 and 9, placed in a small test tube and 0. 5 c.c. etc. Table I). of 5 per cent washed suspension of Brown (1919) suggested that glucose sheep cells was added. The tubes were even in small amount had a marked kept at 370 C. in a water bath and ex- effect on the degree of hemolysis of amined at regular intervals: 20 min., colonies on blood plates.
Load more

Recommended publications
  • Tryptose Blood Agar Base
    Tryptose Blood Agar Base Intended Use Principles of the Procedure Tryptose Blood Agar Base is used with blood in isolating, Tryptose is the source of nitrogen, carbon and amino acids in cultivating and determining the hemolytic reactions of fastidi- Tryptose Blood Agar Base. Beef extract provides additional ous microorganisms. nitrogen. Sodium chloride maintains osmotic balance. Agar is the solidifying agent. Summary and Explanation Investigations of the nutritive properties of tryptose demon- Supplementation with 5-10% blood provides additional growth strated that culture media prepared with this peptone were factors for fastidious microorganisms and is used to determine superior to the meat infusion peptone media previously used hemolytic patterns of bacteria. for the cultivation of Brucella, streptococci, pneumococci, me- Formula ningococci and other fastidious bacteria. Casman1,2 reported Difco™ Tryptose Blood Agar Base that a medium consisting of 2% tryptose, 0.3% beef extract, Approximate Formula* Per Liter 0.5% NaCl, 1.5% agar and 0.03% dextrose equaled fresh beef Tryptose .................................................................... 10.0 g infusion base with respect to growth of organisms. The small Beef Extract ................................................................. 3.0 g amount of carbohydrate was noted to interfere with hemolytic Sodium Chloride ......................................................... 5.0 g Agar ......................................................................... 15.0 g reactions, unless the medium was incubated in an atmosphere *Adjusted and/or supplemented as required to meet performance criteria. of carbon dioxide. Tryptose Blood Agar Base is a nutritious infusion-free basal Directions for Preparation from medium typically supplemented with 5-10% sheep, rabbit or Dehydrated Product horse blood for use in isolating, cultivating and determining 1. Suspend 33 g of the powder in 1 L of purified water.
    [Show full text]
  • Laboratory Exercises in Microbiology: Discovering the Unseen World Through Hands-On Investigation
    City University of New York (CUNY) CUNY Academic Works Open Educational Resources Queensborough Community College 2016 Laboratory Exercises in Microbiology: Discovering the Unseen World Through Hands-On Investigation Joan Petersen CUNY Queensborough Community College Susan McLaughlin CUNY Queensborough Community College How does access to this work benefit ou?y Let us know! More information about this work at: https://academicworks.cuny.edu/qb_oers/16 Discover additional works at: https://academicworks.cuny.edu This work is made publicly available by the City University of New York (CUNY). Contact: [email protected] Laboratory Exercises in Microbiology: Discovering the Unseen World through Hands-On Investigation By Dr. Susan McLaughlin & Dr. Joan Petersen Queensborough Community College Laboratory Exercises in Microbiology: Discovering the Unseen World through Hands-On Investigation Table of Contents Preface………………………………………………………………………………………i Acknowledgments…………………………………………………………………………..ii Microbiology Lab Safety Instructions…………………………………………………...... iii Lab 1. Introduction to Microscopy and Diversity of Cell Types……………………......... 1 Lab 2. Introduction to Aseptic Techniques and Growth Media………………………...... 19 Lab 3. Preparation of Bacterial Smears and Introduction to Staining…………………...... 37 Lab 4. Acid fast and Endospore Staining……………………………………………......... 49 Lab 5. Metabolic Activities of Bacteria…………………………………………….…....... 59 Lab 6. Dichotomous Keys……………………………………………………………......... 77 Lab 7. The Effect of Physical Factors on Microbial Growth……………………………... 85 Lab 8. Chemical Control of Microbial Growth—Disinfectants and Antibiotics…………. 99 Lab 9. The Microbiology of Milk and Food………………………………………………. 111 Lab 10. The Eukaryotes………………………………………………………………........ 123 Lab 11. Clinical Microbiology I; Anaerobic pathogens; Vectors of Infectious Disease….. 141 Lab 12. Clinical Microbiology II—Immunology and the Biolog System………………… 153 Lab 13. Putting it all Together: Case Studies in Microbiology…………………………… 163 Appendix I.
    [Show full text]
  • SHEEP BLOOD AGAR - for in Vitro Use Only - Catalogue No
    SHEEP BLOOD AGAR - For in vitro use only - Catalogue No. PS58 Our Sheep Blood Agar is a highly nutritious Interpretation of Results medium used for the cultivation and isolation of a variety of microorganisms. Sheep Blood Agar can be used as a primary- Our Sheep Blood Agar is based on the Oxoid plating medium. Primary isolation is performed to formulation; the prepared medium is said to offer separate and isolate organisms present in a sample. improved nutritional value resulting in better growth This separation allows for characterization of and larger colony size as well giving more colony types and may indicate the presence of consistent hemolytic reactions especially among the clinically significant bacteria. When examining streptococci. The base is specifically designed for plates a hand lens or stereoscopic microscope use with sheep blood as horse blood has shown to should be available for examining very small give different and conflicting hemolytic reactions colonies. The different types of colonial when incorporated into other blood agars. morphology appearing on the agar plate should be The nutritional components include pancreatic noted as well as the number of each morphotype digest of casein, neutralized peptone, and yeast present. Hemolysis is also a very useful differential extract, and the addition of sodium chloride characteristic that is best viewed when a bright light provides an osmotically balanced medium for is transmitted from behind the plate. Four different bacterial cells. The addition of 5% defibrinated types of hemolysis can be described: sheep blood allows for the determination of hemolytic reactions, an important differential 1. Alpha-hemolysis ( α) – Partial hemolysis that characteristic.
    [Show full text]
  • Screening for Asymptomatic Bacteriuria in Adults: an Updated Systematic Review for the U.S
    Evidence Synthesis Number 183 Screening for Asymptomatic Bacteriuria in Adults: An Updated Systematic Review for the U.S. Preventive Services Task Force Prepared for: Agency for Healthcare Research and Quality U.S. Department of Health and Human Services 5600 Fishers Lane Rockville, MD 20857 www.ahrq.gov Contract No. HHSA-290-2015-00007-I, Task Order No. 3 Prepared by: Kaiser Permanente Research Affiliates Evidence-based Practice Center Kaiser Permanente Center for Health Research Portland, OR Investigators: Jillian T. Henderson, PhD, MPH Elizabeth M. Webber, MS Sarah I. Bean, MPH AHRQ Publication No. 19-05252-EF-1 September 2019 This report is based on research conducted by the Kaiser Permanente Research Affiliates Evidence-based Practice Center (EPC) under contract to the Agency for Healthcare Research and Quality (AHRQ), Rockville, MD (HHSA-290-2015-00007-I, Task Order No. 3). The findings and conclusions in this document are those of the authors, who are responsible for its contents, and do not necessarily represent the views of AHRQ. Therefore, no statement in this report should be construed as an official position of AHRQ or of the U.S. Department of Health and Human Services. The information in this report is intended to help health care decision makers—patients and clinicians, health system leaders, and policymakers, among others—make well-informed decisions and thereby improve the quality of health care services. This report is not intended to be a substitute for the application of clinical judgment. Anyone who makes decisions concerning the provision of clinical care should consider this report in the same way as any medical reference and in conjunction with all other pertinent information (i.e., in the context of available resources and circumstances presented by individual patients).
    [Show full text]
  • Special Microbiology Practical Week 3. – STREPTOCOCCI Streptococci
    Special Microbiology practical week 3. – STREPTOCOCCI Streptococci are Gram-positive, nonmotile, nonsporeforming, catalase-negative cocci that occur in pairs or chains. Older cultures may lose their Gram-positive character. Most streptococci are facultative anaerobes, and some are obligate (strict) anaerobes.Most require enriched media (blood agar). Streptococci are subdivided into groups by antibodies that recognize surface antigens These groups may include one or more species.Serologic grouping is based on antigenic differences in cell wall carbohydrates (groups A to V), in cell wall pili-associated protein, and in the polysaccharide capsule in group B streptococci.Rebecca Lancefield developed the serologic classification scheme in 1933. β-hemolyticstrains possess group- specific cell wall antigens, most of which are carbohydrates. These antigens can be detected by immunologic assays and have been useful for the rapid identification of some important streptococcal pathogens.The most important groupable streptococci are A, B and D. Among the groupable streptococci, infectious disease (particularly pharyngitis) is caused by group A. Group A streptococci have a hyaluronic acid capsule.Streptococcus pneumoniae(a major cause of human pneumonia) and Streptococcus mutansand other so-called viridans streptococci (among the causes of dental caries) do not possess group antigen.Streptococcus pneumoniaehas a polysaccharide capsulethat acts as a virulence factor for the organism,more than 90 different serotypesare known, and these types differ in virulence. Beta Hemolysis Streptococcus pyogenes, or Group A beta-hemolytic Streptococci (GAS), and Streptococcus agalactiae, or Group B beta-hemolytic Streptococci (GBS) blood agar cultures display beta hemolysis. Beta hemolysis (β-hemolysis), called complete hemolysis, is a complete lysisof red blood cells in the media around and under the colonies: the area appears lightened (yellow) and transparent.
    [Show full text]
  • Escherichia Coli Isolates Causing Asymptomatic Bacteriuria in Catheterized and Noncatheterized Individuals Possess Similar Virulence Propertiesᰔ 1 2 1 2
    JOURNAL OF CLINICAL MICROBIOLOGY, July 2010, p. 2449–2458 Vol. 48, No. 7 0095-1137/10/$12.00 doi:10.1128/JCM.01611-09 Copyright © 2010, American Society for Microbiology. All Rights Reserved. Escherichia coli Isolates Causing Asymptomatic Bacteriuria in Catheterized and Noncatheterized Individuals Possess Similar Virulence Propertiesᰔ 1 2 1 2 Rebecca E. Watts, Viktoria Hancock, Cheryl-Lynn Y. Ong, Rebecca Munk Vejborg, Downloaded from Amanda N. Mabbett,1 Makrina Totsika,1 David F. Looke,3 Graeme R. Nimmo,4 Per Klemm,2 and Mark A. Schembri1* School of Chemistry and Molecular Biosciences, University of Queensland, Brisbane, Queensland, Australia1; Microbial Adhesion Group, DTU Food, Technical University of Denmark, Lyngby, Denmark2; and Infection Management Services, Princess Alexandra Hospital,3 and Queensland Health Pathology Service, Department of Microbiology, Royal Brisbane and Women’s Hospital,4 Brisbane, Queensland, Australia Received 19 August 2009/Returned for modification 12 November 2009/Accepted 22 April 2010 http://jcm.asm.org/ Urinary tract infections (UTIs) are among the most common infectious diseases of humans, with Escherichia coli being responsible for >80% of all cases. Asymptomatic bacteriuria (ABU) occurs when bacteria colonize the urinary tract without causing clinical symptoms and can affect both catheterized patients (catheter- associated ABU [CA-ABU]) and noncatheterized patients. Here, we compared the virulence properties of a collection of ABU and CA-ABU nosocomial E. coli isolates in terms of antibiotic resistance, phylogenetic grouping, specific UTI-associated virulence genes, hemagglutination characteristics, and biofilm formation. CA-ABU isolates were similar to ABU isolates with regard to the majority of these characteristics; exceptions were that CA-ABU isolates had a higher prevalence of the polysaccharide capsule marker genes kpsMT II and on October 22, 2015 by University of Queensland Library kpsMT K1, while more ABU strains were capable of mannose-resistant hemagglutination.
    [Show full text]
  • Streptococci
    STREPTOCOCCI Streptococci are Gram-positive, nonmotile, nonsporeforming, catalase-negative cocci that occur in pairs or chains. Older cultures may lose their Gram-positive character. Most streptococci are facultative anaerobes, and some are obligate (strict) anaerobes. Most require enriched media (blood agar). Streptococci are subdivided into groups by antibodies that recognize surface antigens (Fig. 11). These groups may include one or more species. Serologic grouping is based on antigenic differences in cell wall carbohydrates (groups A to V), in cell wall pili-associated protein, and in the polysaccharide capsule in group B streptococci. Rebecca Lancefield developed the serologic classification scheme in 1933. β-hemolytic strains possess group-specific cell wall antigens, most of which are carbohydrates. These antigens can be detected by immunologic assays and have been useful for the rapid identification of some important streptococcal pathogens. The most important groupable streptococci are A, B and D. Among the groupable streptococci, infectious disease (particularly pharyngitis) is caused by group A. Group A streptococci have a hyaluronic acid capsule. Streptococcus pneumoniae (a major cause of human pneumonia) and Streptococcus mutans and other so-called viridans streptococci (among the causes of dental caries) do not possess group antigen. Streptococcus pneumoniae has a polysaccharide capsule that acts as a virulence factor for the organism; more than 90 different serotypes are known, and these types differ in virulence. Fig. 1 Streptococci - clasiffication. Group A streptococci causes: Strep throat - a sore, red throat, sometimes with white spots on the tonsils Scarlet fever - an illness that follows strep throat. It causes a red rash on the body.
    [Show full text]
  • Inhibition of a Staphylococcal Hemolysin by a Soluble Substance Produced by a Nonhemolytic
    INHIBITION OF A STAPHYLOCOCCAL HEMOLYSIN BY A SOLUBLE SUBSTANCE PRODUCED BY A NONHEMOLYTIC MICROCOCCUS SPECIES PINGHUI LIU' Kitasato Institute for Infectious Diseases, Tokyo, Japan Received for publication June 2, 1954 Inhibition of the activity of a bacterial toxin usually grew into the line of micrococcus. The by antitoxic immune sera or by chemicals such as picture resembles those in an article published by formalin is commonly observed. The inhibition Elek and Levy (1950). The inhibition of hemolysis caused by metabolic products of other types of in their cases, however, was caused by specific bacteria, however, has never been described. The antihemolytic sera. In order to avoid the confu- present communication describes a preliminary sion with specific antisera which are often called observation of one such phenomenon. "antihemolysin", the soluble substance produced by the micrococcus will be referred to as "hemol- EXPERIMENTAL METHODS AND RESULTS ysin inhibiting substance" (HIS). A colony of a yellow pigment forming, non- Identification of the hemolysin inhibited by hemolytic Micrococcus species (tentatively identi- hemolysin inhibiting substance. It is well known fied as Micrococcus luteus) was found on a human that staphylococci from various sources produce blood agar plate close to a hemolytic staphylo- several different types of hemolysins (Glenny and coccal colony. It was also noted that the zone of Stevens, 1935; Smith and Price, 1938; Williams hemolysis of this Staphylococcus (the term and Harper, 1947). It was necessary to identify Micrococcus is not used here for this organism to the type of hemolysin concerned in this phe- avoid confusion with the other) was slightly but nomenon.
    [Show full text]
  • Clinical Medical Assistant
    Career Programs Clinical Medical Assistant Clemson University - Center for Corporate Learning 1 North Main Street, 7th Floor, Greenville, SC 29601 http://www.clemson.edu/online/ Contact: Juanita Durham│ 864.656.3984│[email protected] Clinical Medical Assistant Format: Self-Pace Online / eLearning Program Duration: 6 Months Course Contact Hours: 780 The Clinical Medical Assisting Profession The Clinical Medical Assisting program is designed to prepare students to function as professionals in multiple healthcare settings. Medical Assistants with a clinical background perform various clinical tasks including assisting with the administration of medications and with minor procedures, performing an EKG electrocardiogram, obtaining laboratory specimens for testing, educating patients, and other related tasks. Job opportunities are prevalent with physician’s offices, clinics, chiropractor’s offices, hospitals, and outpatient facilities. The Clinical Medical Assisting Program This program prepares students to assist physicians by performing functions related to the clinical aspects of a medical office. Instruction includes preparing patients for examination and treatment, routine laboratory procedures, pharmacology, taking and documenting vital signs, technical aspects of phlebotomy, the 12-lead EKG and the cardiac life cycle. Education and National Certifications • Students should have or be pursuing a high school diploma or GED. • There are no state approval and/or state requirements associated with this program. • National Certification: o
    [Show full text]
  • BD™ Columbia Agar with 5% Sheep Blood
    INSTRUCTIONS FOR USE – READY-TO-USE PLATED MEDIA PA-254005.06 Rev.: Apr 2013 BD Columbia Agar with 5% Sheep Blood INTENDED USE BD Columbia Agar with 5% Sheep Blood is a highly nutritious general purpose medium for the isolation and cultivation of nonfastidious and fastidious microorganisms from clinical specimens. PRINCIPLES AND EXPLANATION OF THE PROCEDURE Microbiological method. Ellner et al.1 in 1966 reported the development of a new blood agar formulation, which has been designated as Columbia Agar. BD Columbia Agar with 5% Sheep Blood derives its superior growth-supporting properties from the combination of two peptones, and yeast extract as a supplier of the B complex vitamins. Corn starch is included to absorb toxic by-products contained in the specimen and serves as an energy source for organisms possessing alpha- amylases. Sheep blood allows detection of hemolytic reactions and supplies the X factor (heme) necessary for the growth of many pathogenic species. On this medium, colonies tend to be larger and growth is more luxuriant than on media containing other blood agar bases. Columbia Blood Agar is recommended as a primary isolation medium in the MiQ standards and in other diagnostic manuals.2,3 In many European countries, this medium has become the most frequently used primary isolation medium for clinical specimens. REAGENTS BD Columbia Agar with 5% Sheep Blood Formula* Per Liter Purified Water Pancreatic Digest of Casein 12.0 g Peptic Digest of Animal Tissue 5.0 Yeast Extract 3.0 Beef Extract 3.0 Corn Starch 1.0 Sodium Chloride 5.0 Agar 13.5 Sheep Blood, Defibrinated 5 % pH 7.3 ± 0.2 *Adjusted and/or supplemented as required to meet performance criteria.
    [Show full text]
  • Staphylococci
    STAPHYLOCOCCI Staphylococci are typical Gram-positive bacteria forming irregular clusters of cocci. Staphylococci are widespread in nature, although they are mainly found on the skin, skin glands and mucous membranes of mammals and birds, but can cause infection under certain circumstances. 1S. aureus is more pathogenic than the other common members of the genus, S. epidermidis and S. saprophyticus. S. epidermidis has been known to cause various hospital-acquired infections (such as prosthetic or indwelling devices), whereas S. saprophyticus is mainly associated with urinary tract infections in young females who are sexually active. Disease processes with S. aureus are numerous. The portal of entry is variable, since they gain access to the body via the skin, the respiratory tract or the genito- urinary tract. Staphylococcus aureus expresses many potential virulence factors: 1. surface proteins - promote colonization of host tissues 2. leukocidin, kinases, hyaluronidase - invasins that promote bacterial spread in tissues 3. capsule, Protein A - surface factors that inhibit phagocytic engulfment 4. carotenoids, catalase - enhance staphylococcal survival in phagocytes 5. protein A, coagulase - immunological disguises 6. hemolysins, leukotoxin, leukocidin - membrane-damaging toxins that lyse eucaryotic cell membranes 7. 2TSST, 3ET - exotoxins that damage host tissues or otherwise provoke symptoms of disease 8. inherent and acquired resistance to antimicrobial agents. Fig. 1 Virulence determinants of Staphylococcus aureus. 1 S. - Staphylococcus 2 TSST - Toxic Shock Syndrome Toxin 3 ET - Exfoliatin Toxin Staphylococci can cause many forms of infection: 1. S. aureus causes superficial skin lesions (boils) and localized abscesses in other sites. 2. S. aureus causes deep-seated infections, such as osteomyelitis and endocarditis and more serious skin infections (furunculosis).
    [Show full text]
  • Preparation of the Blood-Enriched Agar with the Use of Red Cell Suspension
    Vol. 1 No. 1 January 2011 ISSN: 2094-9243 pp. 259-275 Asian Journal of Health International Peer Reviewed Journal Clinical Research Section doi: http://dx.doi.org/10.7828/ajoh.v1i1.168 Preparation of the Blood-Enriched Agar with the Use of Red Cell Suspension CARINA R. MAGBOJOS [email protected] RICHELLE S. ARO MA. CHARISMA S. CARINGAL MELVIN M. CASTILLO DARWIN A. LLANES KAREN D. SUMARAY Lyceum of the Philippines University - Batangas City Date Submitted: April 2, 2009 Plagiarism Detection: Passed Final Revision Complied: June 3, 2010 Flesch Reading Ease: 36.08 Gunning Fog Index: 13.11 Abstract - A culture medium is said to support the growth and development of different microorganisms. Certain bacteria like Staphylococcus aureus and Staphylococcus epidermidis entail hemoglobin found in the red blood cells. Because of its cost effectiveness and availability, expired human blood is being utilized in some developing countries. Despite the widely accepted disadvantages of using human blood as enrichment agent, many laboratories still opt to use it due to the unavailability of sheep blood or due to budgetary reasons. This study determined if the washed expired human blood can be used as an alternative enrichment agent in the preparation of Blood Agar Plate (BAP) culture medium in the isolation of Staphylococcus aureus ATCC 25923 and Staphylococcus epidermidis ATCC 12228. The cultural characteristics and hemolytic reactions of the selected microorganisms were recorded, assessed and compared with their growth in BAP. The stability of the washed expired human blood was evaluated in terms of 259 Asian Journal of Health Clinical Research Section temperature and storage period.
    [Show full text]