This document summarizes various culture methods used to isolate and grow bacteria. It describes streak culture, lawn culture, stab culture, pour plate method, and broth culture. It also discusses different techniques for creating anaerobic conditions, including displacement of oxygen with other gases, chemical/biological methods, and anaerobic chambers. The document provides examples of specific media and methods used for different purposes like isolating pure cultures, antibiotic susceptibility testing, and maintaining stock cultures.
CULTURE METHODS
Indications for culture -
Isolate bacteria in pure cultures.
Demonstrate their properties.
Obtain sufficient growth for preparation of antigens & for other tests.
Typing bacterial isolates.
Antibiotic sensitivity.
Estimate viable counts.
Maintain stock cultures.
Streak culture or surface plating
Lawn or carpet culture
Stroke culture
Stab culture
Pour plate method
Anaerobic methods of culturing bacteria
Streak Culture
Routinely employed for isolation
Platinum / Nichrome loops
LAWN OR CARPET CULTURE
Uniform surface growth
Bacteriophage typing
Antibiotic sensitivity testing
Preparation of bacterial antigens & vaccines
STOKE CULTURE
Tubes containing agar slopes
For slide agglutination & other diagnostic tests.
STAB CULTURE
By puncturing a suitable medium with a long, straight charged wire.
For gelatin liquefaction, stock cultures & motility
POUR PLATE METHOD
1 ml of appropriately diluted inoculum is added to 15 ml of molten agar and poured on petridish.
Colonies appear through out the depth of medium.
Used to estimate viable count, recommended method for quantitative urine cultures.
BROTH CULTURE
Inoculated by a charged loop, pipette or syringes.
For blood cultures & sterility testing.
Bacterial Culture methods and method of anaerobiosisNCRIMS, Meerut
CULTURE METHODS FOR Medical students
Culture methods are done to:
Isolate bacteria in pure culture from the clinical specimens and their idintification by various methods.
Determination of antibiotic sensitivity.
Prepare antigens for serodiagnosis of infective diseases.
Maintain stock cultures.
Methods to isolate the Bacteria
Streak culture
Stroke
Stab
Pour plate
Liquid culture
Special methods for anaerobic cultures
CULTURE METHODS
Indications for culture -
Isolate bacteria in pure cultures.
Demonstrate their properties.
Obtain sufficient growth for preparation of antigens & for other tests.
Typing bacterial isolates.
Antibiotic sensitivity.
Estimate viable counts.
Maintain stock cultures.
Streak culture or surface plating
Lawn or carpet culture
Stroke culture
Stab culture
Pour plate method
Anaerobic methods of culturing bacteria
Streak Culture
Routinely employed for isolation
Platinum / Nichrome loops
LAWN OR CARPET CULTURE
Uniform surface growth
Bacteriophage typing
Antibiotic sensitivity testing
Preparation of bacterial antigens & vaccines
STOKE CULTURE
Tubes containing agar slopes
For slide agglutination & other diagnostic tests.
STAB CULTURE
By puncturing a suitable medium with a long, straight charged wire.
For gelatin liquefaction, stock cultures & motility
POUR PLATE METHOD
1 ml of appropriately diluted inoculum is added to 15 ml of molten agar and poured on petridish.
Colonies appear through out the depth of medium.
Used to estimate viable count, recommended method for quantitative urine cultures.
BROTH CULTURE
Inoculated by a charged loop, pipette or syringes.
For blood cultures & sterility testing.
Bacterial Culture methods and method of anaerobiosisNCRIMS, Meerut
CULTURE METHODS FOR Medical students
Culture methods are done to:
Isolate bacteria in pure culture from the clinical specimens and their idintification by various methods.
Determination of antibiotic sensitivity.
Prepare antigens for serodiagnosis of infective diseases.
Maintain stock cultures.
Methods to isolate the Bacteria
Streak culture
Stroke
Stab
Pour plate
Liquid culture
Special methods for anaerobic cultures
2020 Oregon Wine Symposium | Brushing Up: Current Research and Review of Cell...Oregon Wine Board
Let’s be picky about the quality and quantity of microbes we want in our wine! Join this discussion of current research on effective winery cleaning chemicals, best practices, and practical suggestions for maintaining a clean cellar, as well as identifying cleaning clinch points in winemaking and common winery sanitation needs. A great review for wineries of all shapes and sizes, whether you’re just starting out or wanting to take a closer look at your current cleaning protocols. And of course, if you don’t want to constantly be learning and talking about cleaning, then why be in the wine industry at all?!
recent microbial techniques & advancement in identifying, cultivating,& handl...Karunanidhan3
I tried to include all techniques & diseases that are included in Pharm D 2nd year microbiology syllabus as per PCI. Do suggest if i have to improve my writing skills, on officialkarunanidhan@gmail.com
Production of Enzyme - Lipase.
INTRODUCTION: Lipases are hydrolases capable of catalyzing the hydrolysis of Triglycerols (TAGs) into Glycerol and Fatty acids (FAs).
These enzymes operate at the interfaces of Biphasic systems, which is a phenomenon known as interfacial activation.
These do not require co-factors and are easily immobilized on different matrices.
The active sites of lipases are generally characterized by amino acid triad composed of serine, histidine and aspartate.
Lipases exihibit region-selective properties and enantioselective catalytic behaviour and are considered to be the most versatile catalyst in lipid biotechnology.
These enzymes can be employed in a large number industrial processes ( production of agrochemicals, cosmetics , biodiesel etc.)
HISTORICAL BACKGROUND: In 1856, Claude Bernard first discovered a lipase in pancreatic juice as an enzyme that hydrolyzed insoluble oil droplets and converted them to soluble products.
In 1901, the presence of lipases has been observed for Bacillus prodigiosus , B.pycocyancus and B.fluorescens which represents today’s best studied lipase producing bacteria now named Serratia marcescens , Pseudomonas aeruginosa and P.fluorescens.
Lipase have traditionally been obtained from animal pancreas and are used as a digestive aid for human consumption either in crude mixture with other hydrolases (pancreatin) or as a purified grade.
Lipolase was the first commercial recombinant lipase industialized from the fungus Thermomycesl anugiwnosus and expressed in Aspergillus oryzae in 1994.
PROPERTIES: pH optima
Temperature optima and thermal inactivation
Activation and inactivation of the enzyme
Substrate specificity
SOURCES: Plant lipases:
These have been isolated from the leaves, oils, latex and seeds of oleaginous plants and cereals.
Yeast Lipases:
These include species Candida antartica, Candida rugosa, Candida utilis and Saccharomyces species. The production of Biodiesel includes lipases from Thermomycesl anuginosus.
Animal Lipases:
These include pancreatic and pregastric lipases.
Porcine and Human pancreas were the first sources of lipases used in food processing.
Bacterial Lipases: The genera Pseudomonas and Burkholderia are the most widely used for the production of bacterial lipases. P.aeruginosa produces a cystiene hydrolase solvent tolerant lipase.
Fungal Lipases:
Filamentous fungi are considered to be the best source for production of lipases. The genera includes Aspergillus, Rhizopus , Penicillium , Mucor, Geotrichum and Yarrowia etc.
PRODUCTIONTECHNOLOGY:
UpstreamProcessing:
Screening
Strain selection
Inoculum preparation
Immobilization
Fermentation :
Solid-State Fermentation
Submerged Fermentation
Downstream Processing:
Filtration
Centrifugation
Chromatography
Aqueous two phase
Raw Materials and Nutrients:Olive oil, Palm oil, Coconut oil
wheat Bran, rice bran
yeast extract, peptone
Urea, NaNO2
Sucrose , glucose , fructose
KH2PO4
MgSO4 .7 H2O
Microbial Sources:
Bacillus sp.
Sputum is the liquid substance that is produced from the lower respiratory tract when one coughs.
In addition to mucus, sputum contains many materials that are not visible to the naked eye. It often consists of bacteria, cellular fragments, blood, and pus.
Optimization of ABE Fermentation from Rice Husk Medium using Clostridium acet...pranavdadhich
A chemically defined medium was optimised for the biomass production of Clostridium acetobutylicum in the fermentor using rice husk as the carbon source.
Welcome to TechSoup New Member Orientation and Q&A (May 2024).pdfTechSoup
In this webinar you will learn how your organization can access TechSoup's wide variety of product discount and donation programs. From hardware to software, we'll give you a tour of the tools available to help your nonprofit with productivity, collaboration, financial management, donor tracking, security, and more.
2020 Oregon Wine Symposium | Brushing Up: Current Research and Review of Cell...Oregon Wine Board
Let’s be picky about the quality and quantity of microbes we want in our wine! Join this discussion of current research on effective winery cleaning chemicals, best practices, and practical suggestions for maintaining a clean cellar, as well as identifying cleaning clinch points in winemaking and common winery sanitation needs. A great review for wineries of all shapes and sizes, whether you’re just starting out or wanting to take a closer look at your current cleaning protocols. And of course, if you don’t want to constantly be learning and talking about cleaning, then why be in the wine industry at all?!
recent microbial techniques & advancement in identifying, cultivating,& handl...Karunanidhan3
I tried to include all techniques & diseases that are included in Pharm D 2nd year microbiology syllabus as per PCI. Do suggest if i have to improve my writing skills, on officialkarunanidhan@gmail.com
Production of Enzyme - Lipase.
INTRODUCTION: Lipases are hydrolases capable of catalyzing the hydrolysis of Triglycerols (TAGs) into Glycerol and Fatty acids (FAs).
These enzymes operate at the interfaces of Biphasic systems, which is a phenomenon known as interfacial activation.
These do not require co-factors and are easily immobilized on different matrices.
The active sites of lipases are generally characterized by amino acid triad composed of serine, histidine and aspartate.
Lipases exihibit region-selective properties and enantioselective catalytic behaviour and are considered to be the most versatile catalyst in lipid biotechnology.
These enzymes can be employed in a large number industrial processes ( production of agrochemicals, cosmetics , biodiesel etc.)
HISTORICAL BACKGROUND: In 1856, Claude Bernard first discovered a lipase in pancreatic juice as an enzyme that hydrolyzed insoluble oil droplets and converted them to soluble products.
In 1901, the presence of lipases has been observed for Bacillus prodigiosus , B.pycocyancus and B.fluorescens which represents today’s best studied lipase producing bacteria now named Serratia marcescens , Pseudomonas aeruginosa and P.fluorescens.
Lipase have traditionally been obtained from animal pancreas and are used as a digestive aid for human consumption either in crude mixture with other hydrolases (pancreatin) or as a purified grade.
Lipolase was the first commercial recombinant lipase industialized from the fungus Thermomycesl anugiwnosus and expressed in Aspergillus oryzae in 1994.
PROPERTIES: pH optima
Temperature optima and thermal inactivation
Activation and inactivation of the enzyme
Substrate specificity
SOURCES: Plant lipases:
These have been isolated from the leaves, oils, latex and seeds of oleaginous plants and cereals.
Yeast Lipases:
These include species Candida antartica, Candida rugosa, Candida utilis and Saccharomyces species. The production of Biodiesel includes lipases from Thermomycesl anuginosus.
Animal Lipases:
These include pancreatic and pregastric lipases.
Porcine and Human pancreas were the first sources of lipases used in food processing.
Bacterial Lipases: The genera Pseudomonas and Burkholderia are the most widely used for the production of bacterial lipases. P.aeruginosa produces a cystiene hydrolase solvent tolerant lipase.
Fungal Lipases:
Filamentous fungi are considered to be the best source for production of lipases. The genera includes Aspergillus, Rhizopus , Penicillium , Mucor, Geotrichum and Yarrowia etc.
PRODUCTIONTECHNOLOGY:
UpstreamProcessing:
Screening
Strain selection
Inoculum preparation
Immobilization
Fermentation :
Solid-State Fermentation
Submerged Fermentation
Downstream Processing:
Filtration
Centrifugation
Chromatography
Aqueous two phase
Raw Materials and Nutrients:Olive oil, Palm oil, Coconut oil
wheat Bran, rice bran
yeast extract, peptone
Urea, NaNO2
Sucrose , glucose , fructose
KH2PO4
MgSO4 .7 H2O
Microbial Sources:
Bacillus sp.
Sputum is the liquid substance that is produced from the lower respiratory tract when one coughs.
In addition to mucus, sputum contains many materials that are not visible to the naked eye. It often consists of bacteria, cellular fragments, blood, and pus.
Optimization of ABE Fermentation from Rice Husk Medium using Clostridium acet...pranavdadhich
A chemically defined medium was optimised for the biomass production of Clostridium acetobutylicum in the fermentor using rice husk as the carbon source.
Welcome to TechSoup New Member Orientation and Q&A (May 2024).pdfTechSoup
In this webinar you will learn how your organization can access TechSoup's wide variety of product discount and donation programs. From hardware to software, we'll give you a tour of the tools available to help your nonprofit with productivity, collaboration, financial management, donor tracking, security, and more.
Synthetic Fiber Construction in lab .pptxPavel ( NSTU)
Synthetic fiber production is a fascinating and complex field that blends chemistry, engineering, and environmental science. By understanding these aspects, students can gain a comprehensive view of synthetic fiber production, its impact on society and the environment, and the potential for future innovations. Synthetic fibers play a crucial role in modern society, impacting various aspects of daily life, industry, and the environment. ynthetic fibers are integral to modern life, offering a range of benefits from cost-effectiveness and versatility to innovative applications and performance characteristics. While they pose environmental challenges, ongoing research and development aim to create more sustainable and eco-friendly alternatives. Understanding the importance of synthetic fibers helps in appreciating their role in the economy, industry, and daily life, while also emphasizing the need for sustainable practices and innovation.
Acetabularia Information For Class 9 .docxvaibhavrinwa19
Acetabularia acetabulum is a single-celled green alga that in its vegetative state is morphologically differentiated into a basal rhizoid and an axially elongated stalk, which bears whorls of branching hairs. The single diploid nucleus resides in the rhizoid.
June 3, 2024 Anti-Semitism Letter Sent to MIT President Kornbluth and MIT Cor...Levi Shapiro
Letter from the Congress of the United States regarding Anti-Semitism sent June 3rd to MIT President Sally Kornbluth, MIT Corp Chair, Mark Gorenberg
Dear Dr. Kornbluth and Mr. Gorenberg,
The US House of Representatives is deeply concerned by ongoing and pervasive acts of antisemitic
harassment and intimidation at the Massachusetts Institute of Technology (MIT). Failing to act decisively to ensure a safe learning environment for all students would be a grave dereliction of your responsibilities as President of MIT and Chair of the MIT Corporation.
This Congress will not stand idly by and allow an environment hostile to Jewish students to persist. The House believes that your institution is in violation of Title VI of the Civil Rights Act, and the inability or
unwillingness to rectify this violation through action requires accountability.
Postsecondary education is a unique opportunity for students to learn and have their ideas and beliefs challenged. However, universities receiving hundreds of millions of federal funds annually have denied
students that opportunity and have been hijacked to become venues for the promotion of terrorism, antisemitic harassment and intimidation, unlawful encampments, and in some cases, assaults and riots.
The House of Representatives will not countenance the use of federal funds to indoctrinate students into hateful, antisemitic, anti-American supporters of terrorism. Investigations into campus antisemitism by the Committee on Education and the Workforce and the Committee on Ways and Means have been expanded into a Congress-wide probe across all relevant jurisdictions to address this national crisis. The undersigned Committees will conduct oversight into the use of federal funds at MIT and its learning environment under authorities granted to each Committee.
• The Committee on Education and the Workforce has been investigating your institution since December 7, 2023. The Committee has broad jurisdiction over postsecondary education, including its compliance with Title VI of the Civil Rights Act, campus safety concerns over disruptions to the learning environment, and the awarding of federal student aid under the Higher Education Act.
• The Committee on Oversight and Accountability is investigating the sources of funding and other support flowing to groups espousing pro-Hamas propaganda and engaged in antisemitic harassment and intimidation of students. The Committee on Oversight and Accountability is the principal oversight committee of the US House of Representatives and has broad authority to investigate “any matter” at “any time” under House Rule X.
• The Committee on Ways and Means has been investigating several universities since November 15, 2023, when the Committee held a hearing entitled From Ivory Towers to Dark Corners: Investigating the Nexus Between Antisemitism, Tax-Exempt Universities, and Terror Financing. The Committee followed the hearing with letters to those institutions on January 10, 202
Macroeconomics- Movie Location
This will be used as part of your Personal Professional Portfolio once graded.
Objective:
Prepare a presentation or a paper using research, basic comparative analysis, data organization and application of economic information. You will make an informed assessment of an economic climate outside of the United States to accomplish an entertainment industry objective.
Read| The latest issue of The Challenger is here! We are thrilled to announce that our school paper has qualified for the NATIONAL SCHOOLS PRESS CONFERENCE (NSPC) 2024. Thank you for your unwavering support and trust. Dive into the stories that made us stand out!
The Roman Empire A Historical Colossus.pdfkaushalkr1407
The Roman Empire, a vast and enduring power, stands as one of history's most remarkable civilizations, leaving an indelible imprint on the world. It emerged from the Roman Republic, transitioning into an imperial powerhouse under the leadership of Augustus Caesar in 27 BCE. This transformation marked the beginning of an era defined by unprecedented territorial expansion, architectural marvels, and profound cultural influence.
The empire's roots lie in the city of Rome, founded, according to legend, by Romulus in 753 BCE. Over centuries, Rome evolved from a small settlement to a formidable republic, characterized by a complex political system with elected officials and checks on power. However, internal strife, class conflicts, and military ambitions paved the way for the end of the Republic. Julius Caesar’s dictatorship and subsequent assassination in 44 BCE created a power vacuum, leading to a civil war. Octavian, later Augustus, emerged victorious, heralding the Roman Empire’s birth.
Under Augustus, the empire experienced the Pax Romana, a 200-year period of relative peace and stability. Augustus reformed the military, established efficient administrative systems, and initiated grand construction projects. The empire's borders expanded, encompassing territories from Britain to Egypt and from Spain to the Euphrates. Roman legions, renowned for their discipline and engineering prowess, secured and maintained these vast territories, building roads, fortifications, and cities that facilitated control and integration.
The Roman Empire’s society was hierarchical, with a rigid class system. At the top were the patricians, wealthy elites who held significant political power. Below them were the plebeians, free citizens with limited political influence, and the vast numbers of slaves who formed the backbone of the economy. The family unit was central, governed by the paterfamilias, the male head who held absolute authority.
Culturally, the Romans were eclectic, absorbing and adapting elements from the civilizations they encountered, particularly the Greeks. Roman art, literature, and philosophy reflected this synthesis, creating a rich cultural tapestry. Latin, the Roman language, became the lingua franca of the Western world, influencing numerous modern languages.
Roman architecture and engineering achievements were monumental. They perfected the arch, vault, and dome, constructing enduring structures like the Colosseum, Pantheon, and aqueducts. These engineering marvels not only showcased Roman ingenuity but also served practical purposes, from public entertainment to water supply.
9. Stroke Culture
• Tubes containing agar slopes
• For slide agglutination & other
diagnostic tests.
10. Stab Culture
• By puncturing a
suitable medium
with a long,
straight charged
wire.
• For gelatin
liquefaction, stock
cultures & motility
11. Pour Plate Method
• 1 ml of appropriately
diluted inoculum is added
to 15 ml of molten agar
and poured on petridish.
• Colonies appear through
out the depth of medium.
• Used to estimate viable
count, recommended
method for quantitative
urine cultures.
14. Anaerobic Culture Methods
Anaerobic condition can be achieved by:
• Cultivation in vacuum
• Displacement of oxygen with other gases
• Chemical or biological methods
• By displacement and combustion of oxygen
• By reducing agents
• Anaerobic chamber
16. Chemical or Biological Methods
• Alkaline pyrogallol ( pyrogallic acid in
NaOH) absorbs O2
• Yellow phosphorous
• Rosenthal method - Mixture of
chromium & sulphuric acid
• Gaspak
17. BIOLOGICAL METHODS
Absorption of oxygen from small closed systems has been attempted
by incubation along with
Aerobic bacteria EXAMPLE:- Pseudomonas aeruginosa
Anaerobiosis produced by this method is slow and ineffective.
21. Gaspak
• Method of choice for preparing
anaerobic jars.
• Commercially available as
disposable envelope, containing
chemicals which generate H2 ,
CO2 with the addition of water.
22. Reduction of Oxygen
Using reducing agents:
• 1% glucose
• 0.1% thioglycollate
• 0.1% ascorbic acid
• 0.05% cysteine
23. ANAEROBIC CHAMBER
• It is an anaerobic incubation
system
• It provides oxygen free
atmosphere for inoculating culture
media and for incubation
• It is fitted with airtight rubber
gloves to insert hands for
working with specimens
• The anaerobic chamber contains
catalyst, desiccant, hydrogen gas,
carbon dioxide gas, nitrogen gas
and an indicator
Cultivation in vacuum was attempted by incubating cultures in a vacuum desiccators but it proved to be unsatisfactory. This method is not in use now .
Displacement of oxygen
Displacement of oxygen by inert gases like hydrogen, nitrogen,
carbon dioxide or helium is sometimes employed. Oxygen can never be removed completely by this method. A popular but ineffective method is use of candle
Chemical Methods
Pyrogallol
First introduced by Buchner (1888)
Principle:-Alkaline pyragollol absorbs oxygen
Procedure:- a large tube containing solution of NaOH and pyragollol acid placed inside air tight jar produce an anaerobic conditions
Disadvantages:- small amount of CO is formed during the reaction, may be inhibitory to some bacteria
Chromium and sulphuric acid (Rosenthal Method)
Mixture of chromium and sulphuric acid is used for producing anaerobiosis
Principle:- two chemicals react in presence of oxygen
[O2]
Chromium + Sulphuric acid Chromous + anaerobic
sulphate condition
Alumina pellets coated with palladium- catalyst at room temperature
Reduced methylene blue is used as indicator. Remains colorless anaerobically but turns blue on exposure to O2
Gas pack
Commerically available disposable packet containing pellets of sodium borohydride, cobalt chloride, citric acid and sodium bicarbonate
Principle :- These chemicals generate hydrogen and carbon dioxide when water is added then hydrogen combines with oxygen in the presence of a catalyst
Now this technique is widely used for preparing anaerobic jars
Procedure:- after the inoculated plates are placed inside an air tight jar, the packet of “gas pack “ with water added is kept inside and the lid is tightly closed
Anaerobic broth is an easily prepared anaerobic medium into which pieces of red hot metallic iron are introduced. It is then layered over with sterile vaseline.
Anaerobic broth containing fresh animal tissue, such as rabbit kidney, spleen, testes or heart called Smith-Noguchi medium, supports the growth of many anaerobes.
The most widely employed anaerobic liquid culture are:-
-Thioglycollate broth
-Robertson’s cooked meat medium
Bactec - blood culture method
The sample to be tested is inoculated into one or more vials which are inserted into the BACTEC fluorescent series instrument for incubation and periodic reading.
Each vial contains a chemical sensor which can detect increases in CO2 produced by the growth of microorganisms.
The sensor is monitored by the instrument every ten minutes for an increase in its fluorescence, which is proportional to the amount of CO2 present.
A positive reading indicates the presumptive presence of viable microorganisms in the vial.