IDENTIFICATION OF AEROBIC ACTINOMYCETES

Identification Of Aerobic Actinomycetes-Free PDF

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STATUS OF NATIONAL STANDARD METHODS, National Standard Methods which include standard operating procedures SOPs algorithms and. guidance notes promote high quality practices and help to assure the comparability of diagnostic. information obtained in different laboratories This in turn facilitates standardisation of surveillance. underpinned by research development and audit and promotes public health and patient confidence. in their healthcare services The methods are well referenced and represent a good minimum. standard for clinical and public health microbiology However in using National Standard Methods. laboratories should take account of local requirements and may need to undertake additional. investigations The methods also provide a reference point for method development. National Standard Methods are developed reviewed and updated through an open and wide. consultation process where the views of all participants are considered and the resulting documents. reflect the majority agreement of contributors, Representatives of several professional organisations including those whose logos appear on the. front cover are members of the working groups which develop National Standard Methods Inclusion. of an organisation s logo on the front cover implies support for the objectives and process of preparing. standard methods The representatives participate in the development of the National Standard. Methods but their views are not necessarily those of the entire organisation of which they are a. member The current list of participating organisations can be obtained by emailing. standards hpa org uk, The performance of standard methods depends on the quality of reagents equipment commercial. and in house test procedures Laboratories should ensure that these have been validated and shown. to be fit for purpose Internal and external quality assurance procedures should also be in place. Whereas every care has been taken in the preparation of this publication the Health Protection. Agency or any supporting organisation cannot be responsible for the accuracy of any statement or. representation made or the consequences arising from the use of or alteration to any information. contained in it These procedures are intended solely as a general resource for practising. professionals in the field operating in the UK and specialist advice should be obtained where. necessary If you make any changes to this publication it must be made clear where changes have. been made to the original document The Health Protection Agency HPA should at all times be. acknowledged, The HPA is an independent organisation dedicated to protecting people s health It brings together the. expertise formerly in a number of official organisations More information about the HPA can be found. at www hpa org uk, The HPA aims to be a fully Caldicott compliant organisation It seeks to take every possible.
precaution to prevent unauthorised disclosure of patient details and to ensure that patient related. records are kept under secure conditions1, More details can be found on the website at www evaluations standards org uk Contributions to the. development of the documents can be made by contacting standards hpa org uk. The reader is informed that all taxonomy in this document was correct at time of issue. Please note the references are now formatted using Reference Manager software If you alter or delete text. without Reference Manager installed on your computer the references will not be updated automatically. Suggested citation for this document, Health Protection Agency 2009 Identification of aerobic Actinomycetes National Standard Method. BSOP ID 10 Issue 1 http www hpa standardmethods org uk pdf sops asp. IDENTIFICATION OF AEROBIC ACTINOMYCETES, Issue no 1 Issue date 23 11 09 Issued by Standards Unit Department for Evaluations Standards and Training Page 2 of 15. BSOP ID 10i1, This NSM should be used in conjunction with the series of other NSMs from the Health Protection Agency. www evaluations standards org uk,Email standards hpa org uk.
STATUS OF NATIONAL STANDARD METHODS 2,AMENDMENT PROCEDURE 4. SCOPE OF DOCUMENT 5,INTRODUCTION 5,TECHNICAL INFORMATION LIMITATIONS 9. 1 SAFETY CONSIDERATIONS 10,2 TARGET ORGANISMS 10,3 IDENTIFICATION 10. 3 1 MICROSCOPIC APPEARANCE 10,3 2 PRIMARY ISOLATION MEDIA 11. 3 3 COLONIAL APPEARANCE 11,3 4 TEST PROCEDURES 11, 3 5 THE PRESUMPTIVE IDENTIFICATION OF THE AEROBIC ACTINOMYCETES TO GENUS LEVEL 12.
3 6 FURTHER IDENTIFICATION 12,3 7 STORAGE AND REFERRAL 12. 4 IDENTIFICATION OF GRAM POSITIVE BRANCHING RODS FLOW CHART 12. 5 REPORTING 12,5 1 PRESUMPTIVE IDENTIFICATION 12,5 2 CONFIRMATION OF IDENTIFICATION 12. 5 3 MEDICAL MICROBIOLOGIST 12,5 4 CCDC 12,5 5 CENTRE FOR INFECTIONS 12. 5 6 INFECTION CONTROL STAFF 12,6 REFERRALS 12,6 1 REFERENCE LABORATORY 12. 7 ACKNOWLEDGEMENTS AND CONTACTS 13,REFERENCES 14,IDENTIFICATION OF AEROBIC ACTINOMYCETES.
Issue no 1 Issue date 23 11 09 Issued by Standards Unit Department for Evaluations Standards and Training Page 3 of 15. BSOP ID 10i1, This NSM should be used in conjunction with the series of other NSMs from the Health Protection Agency. www evaluations standards org uk,Email standards hpa org uk. AMENDMENT PROCEDURE,Controlled document BSOP ID 10. Controlled document title Identification Of Aerobic Actinomycetes. Each National Standard Method has an individual record of amendments The current amendments. are listed on this page The amendment history is available from standards hpa org uk. On issue of revised or new pages each controlled document should be updated by the copyholder in. the laboratory, Amendment Issue no Insert Page Section s involved Amendment. Number Discarded Issue,IDENTIFICATION OF AEROBIC ACTINOMYCETES.
Issue no 1 Issue date 23 11 09 Issued by Standards Unit Department for Evaluations Standards and Training Page 4 of 15. BSOP ID 10i1, This NSM should be used in conjunction with the series of other NSMs from the Health Protection Agency. www evaluations standards org uk,Email standards hpa org uk. IDENTIFICATION OF AEROBIC,ACTINOMYCETES SPECIES,SCOPE OF DOCUMENT. This document describes the identification of branching Gram positive bacilli isolated from clinical. specimens Colonies may be isolated on blood agar or egg containing media. Anaerobic sporing organisms are described in BSOPID 8 Identification of Clostridium species. Anaerobic cocci are described in BSOP ID 14 Identification of anaerobic cocci Anaerobic Gram. negative rods are described in BSOP ID 25 Identification of anaerobic Gram negative rods. INTRODUCTION, The nomenclature of the group comprising the branching Gram positive rods is complicated. Considerable morphological diversity is not only seen among genera but also among strains of the. same taxon,Characteristics2,Nocardia species, Nocardia species produce rudimentary to extensively branched vegetative hyphae 0 5 1 2 m in.
diameter which grow on the surface and penetrate agar media The hyphae often fragment into rod. shaped or coccoid elements Aerial hyphae are almost always produced Short to long chains of. conidia may be found on the aerial hyphae and occasionally on substrate hyphae Cells stain Gram. positive to Gram variable and are usually acid fast Growth is aerobic producing chalky matt or. velvety colonies Colonial morphology will vary according to the medium or incubation temperature. used The colonies may be brown tan pink orange red purple grey or white Colonies on solid. media may be smooth and moist or granular irregular wrinkled or heaped with a velvety surface due. to aerial filamentation Soluble brown or yellow pigments may be produced Nocardia are catalase. positive and grow on Sabouraud s glucose agar blood agar brain heart infusion agar and. Lowenstein Jensen medium Added carbon dioxide 10 promotes more rapid growth On. Sabouraud dextrose agar colonies of N asteroides complex vary from salmon pink to orange. N brasiliensis colonies are usually orange tan N otitidiscavarum colonies are pale tan whereas. N transvalensis may vary in colour from pale tan to violet Colonies in pure culture can grow after. only 48 hours incubation In mixed cultures other rapidly growing bacteria may obscure small. Nocardia species colonies which may take several weeks to develop Modified Thayer Martin medium. or buffered charcoal yeast extract agar may enhance recovery of Nocardia species3. Microscopic examination of Gram stained clinical specimens may give a rapid and specific diagnosis. Thin delicate weakly to strongly Gram positive irregularly stained or beaded branching filaments are. characteristic of Nocardia species Multiple clinical specimens should be submitted for culture. Nocardia species may not be detected unless pus from a discharging fistula or abscess is examined. Smears and cultures of specimens are often negative unless specimens are obtained by biopsy. Routine blood cultures are not usually positive Many Nocardia species from clinical material are. variably acid fast on primary isolation This is rapidly lost in subcultured colonies Modified Kinyoun. stain decolourised with a weak acid 1 2 sulphuric acid instead of acid alcohol should be used A. single Nocardia colony isolated from CSF or a normally sterile site such as soft tissue abscess pleural. space or joint fluid from a patient with an appropriate clinical presentation should never be ignored. These organisms are seldom laboratory contaminants and are not part of the body s normal flora. Sputum digestion procedures eg N acetyl L cysteine or sodium hydroxide may produce negative. Nocardia species cultures There are currently no serodiagnostic tests available4. IDENTIFICATION OF AEROBIC ACTINOMYCETES, Issue no 1 Issue date 23 11 09 Issued by Standards Unit Department for Evaluations Standards and Training Page 5 of 15. BSOP ID 10i1, This NSM should be used in conjunction with the series of other NSMs from the Health Protection Agency. www evaluations standards org uk,Email standards hpa org uk. Since Nocardia species are ubiquitous in nature the isolation of these microorganisms from. specimens may not be significant clinically The presence of Nocardia in sputum culture may not. always indicate invasive infection but may reflect laboratory contamination or respiratory colonization. The clinical and microbiological difficulties include the non specific presentation of the infection a. frequent requirement for invasive diagnostic biopsy procedures difficulty in isolating the Nocardia and. problems in identification and taxonomic classification N farcinica is commonly misidentified as. N asteroides or Rhodococcus or Gordona species,Streptomyces species. Streptomyces species produce vegetative hyphae 0 5 2 0 m in diameter which form an extensively. branched mycelium which rarely fragments This matures to form chains of three to many non motile. spores A few species produce spores on the substrate mycelium Cells are Gram positive but not. acid alcohol fast Growth is obligately aerobic and the optimum growth temperature is 25 C 35 C. Initially the colonies produced are relatively smooth surfaced but later they develop aerial mycelium. which may appear floccose granular powdery or velvety Colonies are discrete lichenoid leathery or. butyrous The vegetative and aerial mycelia may be pigmented and diffusible pigments may also be. produced Metabolism is oxidative and the catalase test is positive Nitrates are reduced to nitrites. and aesculin is degraded,Rhodococcus species, Rhodococcus species produce cocci which may germinate into short rods form filaments with side.
projections branching or extensively branched hyphae The next generation of cocci or short rods is. produced by fragmentation of the rods filaments or hyphae Microscopic aerial hyphae and spores. are not usually produced and spores are not produced Cells stain Gram positive and are usually. partially acid fast Growth occurs aerobically, There are three main colony types of R equi The classic colony type is pale pink and slimy The. second is colonet and non slimy and the third is pale yellow non slimy and more opaque Colonies. of other rhodococci may be rough smooth or mucoid and pigmented cream buff yellow coral. orange or red Colourless variants may occur particularly of R equi R equi has a variable. microscopic morphology bacillary to coccoid forms and may be discarded as a contaminiant5 The. cyclic variation in morphology of R equi and some non equi rhodococci depends upon incubation time. and growth conditions All rhodococci from clinical specimens are weakly acid fast Colonial and cell. morphology cannot be used distinguish among Rhodococcus Gordonia and Tsukamurella species. Commercial identification systems do not provide reliable identification of Rhodococcus species and. clinically important isolates should be referred to the Reference Laboratory6. Oerskovia species, Oerskovia species produce extensively branching vegetative hyphae approximately 0 5 m in. diameter which grow on the surface and penetrate into agar The hyphae break up into rod shaped. motile flagellate rods Non motile strains may also occur An aerial mycelium is not formed Cells. stain Gram positive although part of the thallus may become Gram negative with age and. coryneforms may be seen Growth is facultatively anaerobic and the catalase test is positive when. IDENTIFICATION OF AEROBIC ACTINOMYCETES Issue no 1 Issue date 23 11 09 Issued by Standards Unit Department for Evaluations Standards and Training Page 1 of 15 BSOP ID 10i1 This NSM should be used in conjunction with the series of other NSMs from the Health Protection Agency www evaluations standards org uk Email standards hpa org uk G3 NATIONAL STANDARD METHOD IDENTIFICATION OF AEROBIC

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