Satellitism Test for the Identification of Haemophilus influenzae: Introduction, Principle, Test Requirements, Procedure, Result -Interpretation, and Keynotes

Introduction

Satellitism test or satellite test is applied to identify Haemophilus influenzae. It is a fastidious bacterium and thus it needs extra ingredients like hemin (factor X) and  NAD (factor V) to grow. It uses the X factor to produce essential respiratory enzymes such as cytochromes, catalases, and peroxidase whereas the V factor is an electron carrier in the organism’s oxidation-reduction system.

Satellitism Test for the Identification of Haemophilus influenzae- Introduction, Principle, Test Requirements, Procedure, Result -Interpretation and Keynotes
Fig. Satellitism Test Positive- Haemophilus influenzae on blood agar

Principle of Satellitism Test

Haemophilus influenaze requires both X and V factors for multiplication. Most strains of Haemophilus species do not grow on 5% sheep blood agar, which contains the X factor but lacks the V factor. Staphylococcus aureus produces the V factor as a metabolic by-product when growing in a bacteria culture containing blood. Therefore,  Haemophilus species may grow on sheep blood agar (BAP) very close to the colonies of S. aureus (as V factor); this phenomenon is known as satelliting, and the test to detect it is called satellitism test.

Test Requirements for Satellitism Test

  • 5% sheep blood agar (BAP) plate
  • Physiological saline/ peptone water
  • Nutrient agar or  tryptic soy agar
  • Suspected growth of H. influenzae ( any organism growing on chocolate agar  and not blood agar that shows in Gram stain, gram-negative coccobacilli or short rods)
  • Bunsen burner
  • Inoculating loop
  • Sterile swab sticks
  • S. aureus (ATCC 25923)
  • CO2 Incubator
  • Control strains (H. influenaze  ATCC 43065 and S. aureus ATCC 25923 )

Test Procedure

  1. First put on gloves
  2. Mix a loopful of suspected colonies of Haemophilus colonies ( test organism) in about 2 ml of sterile physiological saline or sterile peptone water.
  3. Make sure none of the chocolate agar media is transferred.
  4. Using a sterile swab stick, inoculate the test organism suspension on a plate of nutrient agar or tryptic soy agar and a plate of blood agar and spread uniformly.
  5. Streak a pure culture of S. aureus across each of the inoculated plates.
  6. Incubate both plates in a CO2-enriched atmosphere at 35 to 37°C for 18-24 hours.
  7. Observe the culture plates for growth and satellite colonies.

Observation

Observe the growth of organisms on inoculated agar plates like nutrient agar or tryptic soy agar and blood agar.

Result- Interpretations of Satellitism Test 

  1. The suspected colonies are of Haemophilus influenzae if growth is seen in the blood agar but not in the nutrient agar or tryptic soy agar plate.
  2. The colonies near the column of Staphylococcus aureus growth are larger than those furthest from it.
  3. If satellite colonies are present on both blood and nutrient agar plates then the organism is probably a Haemophilus species that requires only factor V, such as Haemophilus parainfluenzae.

Keynotes on Satellitism Test 

  1. The satellitism assay is performed on blood agar and blood agar contains only X factor and V factor is produced by growing S. aureus.
  2. Satellitism is also shown by variant streptococci like Abiotrophia defectiva and Granulicatella species. They do not grow on blood agar, as it lacks nutritional requirements, but will grow in the medium surrounding staphylococci, which supplies the needed nutrients. In Gram stain, these organisms are cocci, filaments, and bulbous forms in smear.
  3. Haemophilus  influenzae is the first free-living organism whose complete genome has been sequenced.
  4. H.  influenzae strains have been classified into 6 capsular types (a-f) based on polysaccharide capsules.
  5.  Among them, only type b is responsible for 95% of the disease.
  6. Some microorganisms grow only on chocolate agar and will not grow on blood agar even with a staphylococcus dot. These organisms include Haemophilus ducreyi, Francisella tularensis, and some Methylobacterium species.

Further Readings

  1. Cowan & Steel’s Manual for identification of Medical Bacteria. Editors: G.I. Barron & R.K. Felthani, 3rd ed 1993, Publisher Cambridge University Press.
  2. Clinical Microbiology Procedure Handbook, Chief in editor H.D. Isenberg, Albert Einstein College of Medicine, New York, Publisher ASM (American Society for Microbiology), Washington DC.
  3. Colour Atlas and Textbook of Diagnostic Microbiology. Editors: Koneman E.W., Allen D.D., Dowell V.R. Jr, and Sommers H.M.
  4. Bailey & Scott’s Diagnostic Microbiology. Editors: Bettey A. Forbes, Daniel F. Sahm & Alice S. Weissfeld, 12th ed 2007, Publisher Elsevier.
  5. Jawetz, Melnick and Adelberg’s Medical Microbiology. Editors: Geo. F. Brook, Janet S. Butel & Stephen A. Morse, 21st ed 1998, Publisher Appleton & Lance, Co Stamford Connecticut.
  6. Mackie and Mc Cartney Practical Medical Microbiology. Editors: J.G. Colle, A.G. Fraser, B.P. Marmion, A. Simmous, 4th ed, Publisher Churchill Living Stone, New York, Melborne, Sans Franscisco 1996.
  7.  Textbook of Diagnostic Microbiology. Editors: Connie R. Mahon, Donald G. Lehman & George Manuselis, 3rd edition2007, Publisher Elsevier.
  8. https://onlinelibrary.wiley.com/doi/pdf/10.1046/j.1365-2958

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    總之,2024年的娛樂城呈現出一個高度融合了技術、安全和負責任賭博的行業新面貌,為玩家提供了前所未有的娛樂體驗。隨著這些趨勢的持續發展,我們可以預見,娛樂城將不斷地創新和進步,為玩家帶來更多精彩和安全的娛樂選擇。

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  65. 2024娛樂城的創新趨勢

    隨著2024年的到來,娛樂城業界正經歷著一場革命性的變遷。這一年,娛樂城不僅僅是賭博和娛樂的代名詞,更成為了科技創新和用戶體驗的集大成者。

    首先,2024年的娛樂城極大地融合了最新的技術。增強現實(AR)和虛擬現實(VR)技術的引入,為玩家提供了沉浸式的賭博體驗。這種全新的遊戲方式不僅帶來視覺上的震撼,還為玩家創造了一種置身於真實賭場的感覺,而實際上他們可能只是坐在家中的沙發上。

    其次,人工智能(AI)在娛樂城中的應用也達到了新高度。AI技術不僅用於增強遊戲的公平性和透明度,還在個性化玩家體驗方面發揮著重要作用。從個性化遊戲推薦到智能客服,AI的應用使得娛樂城更能滿足玩家的個別需求。

    此外,線上娛樂城的安全性和隱私保護也獲得了顯著加強。隨著技術的進步,更加先進的加密技術和安全措施被用來保護玩家的資料和交易,從而確保一個安全可靠的遊戲環境。

    2024年的娛樂城還強調負責任的賭博。許多平台採用了各種工具和資源來幫助玩家控制他們的賭博行為,如設置賭注限制、自我排除措施等,體現了對可持續賭博的承諾。

    總之,2024年的娛樂城呈現出一個高度融合了技術、安全和負責任賭博的行業新面貌,為玩家提供了前所未有的娛樂體驗。隨著這些趨勢的持續發展,我們可以預見,娛樂城將不斷地創新和進步,為玩家帶來更多精彩和安全的娛樂選擇。

    Reply
  66. Деревянные дома под ключ
    Дома АВС – Ваш уютный уголок

    Мы строим не просто дома, мы создаем пространство, где каждый уголок будет наполнен комфортом и радостью жизни. Наш приоритет – не просто предоставить место для проживания, а создать настоящий дом, где вы будете чувствовать себя счастливыми и уютно.

    В нашем информационном разделе “ПРОЕКТЫ” вы всегда найдете вдохновение и новые идеи для строительства вашего будущего дома. Мы постоянно работаем над тем, чтобы предложить вам самые инновационные и стильные проекты.

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    Для тех, кто ценит экологичность и близость к природе, мы предлагаем деревянные дома премиум-класса. Используя клееный брус и оцилиндрованное бревно, мы создаем уникальные и здоровые условия для вашего проживания.

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    С Домами АВС создайте свой уютный уголок, где каждый момент жизни будет наполнен радостью и удовлетворением

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  67. Дома АВС – Ваш уютный уголок

    Мы строим не просто дома, мы создаем пространство, где каждый уголок будет наполнен комфортом и радостью жизни. Наш приоритет – не просто предоставить место для проживания, а создать настоящий дом, где вы будете чувствовать себя счастливыми и уютно.

    В нашем информационном разделе “ПРОЕКТЫ” вы всегда найдете вдохновение и новые идеи для строительства вашего будущего дома. Мы постоянно работаем над тем, чтобы предложить вам самые инновационные и стильные проекты.

    Мы убеждены, что основа хорошего дома – это его дизайн. Поэтому мы предоставляем услуги опытных дизайнеров-архитекторов, которые помогут вам воплотить все ваши идеи в жизнь. Наши архитекторы и персональные консультанты всегда готовы поделиться своим опытом и предложить функциональные и комфортные решения для вашего будущего дома.

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    Для тех, кто ценит экологичность и близость к природе, мы предлагаем деревянные дома премиум-класса. Используя клееный брус и оцилиндрованное бревно, мы создаем уникальные и здоровые условия для вашего проживания.

    Тем, кто предпочитает надежность и многообразие форм, мы предлагаем дома из камня, блоков и кирпичной кладки.

    Для практичных и ценящих свое время людей у нас есть быстровозводимые каркасные дома и эконом-класса. Эти решения обеспечат вас комфортным проживанием в кратчайшие сроки.

    С Домами АВС создайте свой уютный уголок, где каждый момент жизни будет наполнен радостью и удовлетворением

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  68. online pay per click
    Understanding the processes and protocols within a Professional Tenure Committee (PTC) is crucial for faculty members. This Frequently Asked Questions (FAQ) guide aims to address common queries related to PTC procedures, voting, and membership.

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    Vote justification forms provide transparency in decision-making. Members articulate their reasoning, fostering a culture of openness and ensuring that decisions are well-founded and understood by the academic community.

    2. How can absentee ballots be cast?
    To accommodate absentee voting, PTCs may implement secure electronic methods or designated proxy voters. This ensures that faculty members who cannot physically attend meetings can still contribute to decision-making processes.

    3. How will additional members of PTCs be elected in departments with fewer than four tenured faculty members?
    In smaller departments, creative solutions like rotating roles or involving faculty from related disciplines can be explored. Flexibility in election procedures ensures representation even in departments with fewer tenured faculty members.

    4. Can a faculty member on OCSA or FML serve on a PTC?
    Faculty members involved in other committees like the Organization of Committee on Student Affairs (OCSA) or Family and Medical Leave (FML) can serve on a PTC, but potential conflicts of interest should be carefully considered and managed.

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    A positive vote typically indicates approval or agreement, while a negative vote signifies disapproval or disagreement. Clear definitions and guidelines within each PTC help members interpret and cast their votes accurately.

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    A quorum, the minimum number of members required for a valid meeting, is essential for decision-making. Specific rules about quorum size are usually outlined in the PTC’s governing documents.

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