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The addition of a trapping agent (Gram's iodine) rapid decolorization with alcohol or acetone, and;Escherichia coli ATCC ® Stains blue in color Staphylococcus aureus ATCC ® Stains blue in color Corynebacterium diphtheriae ATCC ® 8028 Cell appears banded or beaded with deep blue granules and a lighter blue cytoplasmCounterstaining with safranin or carbol fuchsin (it will more intensely stain anaerobic bacteria)
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E.coli gram stain morphology and arrangement
E.coli gram stain morphology and arrangement-Gramstain Grampositive cocci Microscopic appearance Cocci in clusters, short chains, diplococci and single cocci Clinical significance Enterococcus faecalisis a Grampositive, commensal bacterium inhabiting the gastrointestinal tracts of humans and other mammalsE Coli Microscopy To determine whether a strain (s) is present in a sample, it's necessary to stain the sample Here, Gram stain is used as it helps distinguish between the gram positive and gram negative bacteria in a sample Being a differential stain, Gram stain is more complex compared to more simple stains like methylene blue
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The colony morphology of organisms is observed Colony shape, size (in mm), color, consistency, elevation, opacity, and margin are observed and noted down for further identification Gram staining for identifying gram negative bacteria Gram staining is the beginning test in an identification procedure in bacterial classificationStart studying Morphology of bacteria and gram stain Learn vocabulary, terms, and more with flashcards, games, and other study tools ( Gram ve Bacilli) Ecoli Identify the bacteria (Gram ve Bacilli) Pseudomonas aeruginosa ( Decolonization) gram negative will lose the stain Process D in both Safranin Process E in bothPercentage of isolation of the five E coli Colonial Morphology (CM) classes from inpatients and outpatients (EMB) agar, also the result of Gram staining agrees with the findings of the study
Gram Staining reaction – Gram ve uniform turbidity is produced which is further analyzed for the morphology (under the microscope), gram reaction, biochemical tests, and staphylococcus specific tests That's all about the Morphology & Cultural Characteristics of Staphylococcus aureusGram positive bacteria stain bluepurple and Gram negative bacteria stain red The difference between the two groups is believed to be due to a much larger peptidoglycan (cell wall) in Gram positivesCharacteristic fusiform morphology (Gram stain) Capnocytophaga ochracea Gramnegative rods from a culture showing characteristic fusiform morphology (Gram stain) Escherichia coli Atypical growth in a blood culture from a patient under antibiotic treatment Very long threads, some poorly stained (Gram stain)
Through this experiment, gram staining skills develop More understanding the types and morphology of bacteria Expected experimental result, Escherichia coli (Ecoli ) is a negative gram bacteria which stain pink colour , while Staphylococcus aureus ( Saureus ) is a positive gram bacteria which stain purple colour Materials Bacteria Escherichia coli , Staphylococcus aureus Crystal Violet Gram's iodine Absolute alcohol Safranin Methodology As per manualThe Gram stain is used to differenciate Grampositive from Gramnegative bacteria Grampositive bacteria have a thicker peptidoglycan layer and therefore retain the primary stain (crystal violet) whereas Gramnegative cells lose it when treated with a decolourizer (absolute alcohol) They then take in the secondary stain (iodine)On Endo agar it looks like lactose negative)All four strains are mannitol positive (best seen in fig D), cellobiose negative (strains A, B)
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Positive Gram Stain Negative E coli Serratia marcescens Proteus mirabilis Neisseria gonorrhea Alcaligenes Faecalis Proteus vulgaris Klebsiella pneumoniae Citrobacter freundii Pseudomonas fluorescens Enterobacter aerogenes Staphylococcus aureus Streptococcus pneumoniae Bacillus cereus Enterococcus faecalis Micrococcus luteus E coli SerratiaGram staining technique is a fourstep procedures used to distinguish bacteria into two large groups, Grampositive and Gramnegative The reagents used are crystal violet, iodine, 95% alcohol and safranin and they are applied sequentially The bacteria used are Micrococcus luteus and Escherichia coliThe Gram stain is the most widely used staining procedure in bacteriology It is called a differential stain since it differentiates between Grampositive and Gramnegative bacteria Bacteria that stain purple with the Gram staining procedure are termed Grampositive;
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Gram stain morphology for EscherichiaEscherichia coli Gramnegative Short rods (bacilli) Encapsulated and Unencapsulated Gram stain morphology for FrancisellaFrancisella tularensisCommon examples of gramnegative include Salmonella spp, Escherichia coli (E coli) and the Enterobacteriaceae spp GramPositive Bacteria Unlike gramnegative, gram positive bacteria have a thick peptidoglycan layer that allows them to retain the primary stain/dye (crystal violet stain)Cells are typically rodshaped, and are about μm long and 025–10 μm in diameter, with a cell volume of 06–07 μm 3 E coli stains Gramnegative because its cell wall is composed of a thin peptidoglycan layer and an outer membrane During the staining process, E coli picks up the color of the counterstain safranin and stains pink
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Start studying Morphology of bacteria and gram stain Learn vocabulary, terms, and more with flashcards, games, and other study tools ( Gram ve Bacilli) Ecoli Identify the bacteria (Gram ve Bacilli) Pseudomonas aeruginosa ( Decolonization) gram negative will lose the stain Process D in both Safranin Process E in bothThe Gram stain is a differential staining technique used to classify & categorize bacteria into two major groups Gram positive and Gram negative, based on the differences of the chemical and physical properties of the cell wall Escherichia coli Fig Gram negative bacteria Bacterial Morphology Bacteria are very small unicellularEscherichia coli Four different strains of Escherichia coli on Endo agar with biochemical slope Glucose fermentation with gas production, urea and H 2 S negative, lactose positive (with exception of strain D "late lactose fermenter";
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On staining, E coli appear as nonsporeforming, Gramnegative rodshaped bacterium;The genus Escherichia is named after Theodor Escherich, who isolated the type species of the genusEscherichia organisms are gramnegative bacilli that exist singly or in pairsE coli isTHE PROCEDURE done individually We have cultures of E coli and Bacillus for you to gram stainThis will give you gram and gram – controls to check your procedure against You can use 2 slides, 1 for each bacterium, or you can divide one slide in half and smear each bacterium on the divided slide
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We have cultures of E coli and Bacillus for you to gram stain This will give you gram and gram – controls to check your procedure against You can use 2 slides, 1 for each bacterium, or you can divide one slide in half and smear each bacterium on the divided slideGram Staining of Micrococcus luteus,Escherichia coli, and Serratia marcescens INTRODUCTION In 14, a Danish botanist named Hans Christian Gram developed the popular staining technique known as the "gram stain"His background in studying plants at the University of Copenhagen introduced him to observing different tissues under a microscope, and thus propelled his interest in pharmacologyE coli are Gramnegative bacteria, meaning that they do not retain the crystal violet stain commonly used to differentiate bacteria Their status as Gramnegative bacteria is due to their thin cell walls E coli has cell walls made out of two thing peptidoglycan layers, an inner and outer membrane
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Escherichia coli or in short E coli is a rodshaped microorganism It is categorized as gramnegative based on the gram staining procedure E coli is usually found in the large intestine of mammals and is the most abundant microorganism to be present in the feces Hence, it is used as an indicator for water purityYour unknown sample For example, when you perform a Gram stain, you will always include samples of Staphylococcus epidermidis (S epi), which is known to be Gram positive, and Escherichia coli, which is known to be Gramnegative If the Gram stain procedure works as it should, S epi will be purple and E coli will be pinkThe result shows that the smear named S aureus is a Grampositive organism because it shows a purple in colour while the E coli is Gram negative organism because it shows a pinkred in colour Conclusion From the experiment, the gram staining is the method to distinguish and differentiate between Grampositive bacteria and Gramnegative bacteria
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E coli in gram stain E coli in Gram stain showing gram negative rods having size of about µm long and diameter as shown above picture Introduction of Escherichia coli E coli is a Gram negative, aerobe and facultative anaerobic, rodshaped bacteria Optimal temperature for growth is 3617°C with most strains growing over the range 1844 °CPositive Gram Stain Negative E coli Serratia marcescens Proteus mirabilis Neisseria gonorrhea Alcaligenes Faecalis Proteus vulgaris Klebsiella pneumoniae Citrobacter freundii Pseudomonas fluorescens Enterobacter aerogenes Staphylococcus aureus Streptococcus pneumoniae Bacillus cereus Enterococcus faecalis Micrococcus luteus E coli SerratiaIn Gramnegative bacteria, the cell wall is only 1–3 layers thick , and in E coli 80% or more of the peptidoglycan exists as a monolayer Consistent with these earlier results, recent electron cryotomography density profile measurements have revealed that the thickness of the cell wall of both E coli and another Gramnegative bacteria Caulobacter crescentus is at most 4 nm ( 13 )
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Microorganisms are very small microscopic structures that are capable of free living Some of the microorganisms are nonpathogenic and live on the body of human beings ie on the skin, in the nostrils, in the intestinal tract etc, and they are called commensals The organisms that are capable of causing disease are called pathogenic organisms These are two groups depending upon theE coli S epidermidis Note Escherichia coli is a tiny pink (Gram) rod Staphylococcus epidermidis is a purple (Gram) sphere or coccus Draw a picture of a typical microscopic field and identify both Escherichia coli and Staphylococcus epidermidis Record this in the results section for this lab Colored pencils are available throughout the roomProcedure of Gram stain for E coli Cover the smear with crystal violet and allow it to stand for one minute Rinse the smear gently under tap water Cover the smear with Gram's iodine and allow it to stand for one minute Rinse smear again gently under tap water Decolorize the smear with 95% alcohol Rinse the smear again gently under tap
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Percentage of isolation of the five E coli Colonial Morphology (CM) classes from inpatients and outpatients (EMB) agar, also the result of Gram staining agrees with the findings of the studyGram stain morphology for EnterococcusEnterococcus faecalis Enterococcus faecium Grampositive Round to ovoid (cocci) pairs or chains Gram stain morphology for EscherichiaEscherichia coli Gramnegative Short rods (bacilli) Encapsulated and UnencapsulatedGram Positive vs Gram Negative Bacteria This is a mixed culture of Gram negative Escherichia coli (redorange) and Gram positive Staphylococcus aureus (bluepurple) stained using the Gram staining method Michael R Francisco/Flickr/CC BYSA
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Routine urine cultures should be plated using calibrated loops for the semiquantitative method Note The most commonly used criterion for defining significant bacteriuria is the presence of ⩾10 5 CFU per milliliter of urine2 Morphology and Staining of Escherichia Coli E coli is Gramnegative straight rod, 13 µ x 0407 µ, arranged singly or in pairs (Fig 281) It is motile by peritrichous flagellae, though some strains are nonmotile Spores are not formed Capsules and fimbriae are found in some strains 3 Cultural Characteristics of Escherichia ColiThose that stain pink are said to be Gramnegative The terms positive and negative have nothing to do with electrical charge, but simply designate two distinct morphological groups of bacteria
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Gramstain Grampositive cocci Microscopic appearance Cocci in clusters, short chains, diplococci and single cocci Clinical significance Enterococcus faecalisis a Grampositive, commensal bacterium inhabiting the gastrointestinal tracts of humans and other mammalsIts cell wall contains mycolic acids, long, branched fatty acids that are normally present in scidfast bacteria The acids prevent proper gram staining that would normally identify the cell as a gram positive cell because they create a waxy coating so the crystal violet has difficulty entering the cell, therefore making it seem gramnegativeGram negative are the type of bacteria that do not retain the primary stain During decolorization, these bacteria lose the crystal violet stain (primary stain) because they have a thin Peptidoglycan layer However, they take up the counter stain (safranin) and will appear reddish or pink when viewed under the microscope
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Escherichia coli O157H7 was first identified as a human pathogen in 19 in the United States of America, following an outbreak of bloody diarrhea associated with contaminated hamburger meat In 06, there was an outbreak involving raw spinach, with 199 illnesses, 102 hospitalizations, 31 hemolytic uremic syndrome, a severe kidney conditionProcedure of Gram stain for E coli Cover the smear with crystal violet and allow it to stand for one minute Rinse the smear gently under tap water Cover the smear with Gram's iodine and allow it to stand for one minute Rinse smear again gently under tap water Decolorize the smear with 95% alcohol Rinse the smear again gently under tap waterInvestigative procedure in biology A Gram stain of mixed Staphylococcus aureus ( S aureus ATCC , Grampositive cocci, in purple) and Escherichia coli ( E coli ATCC , gramnegative bacilli, in red), the most common Gram stain reference bacteria Gram stain or Gram staining, also called Gram's method, is a method of staining used to distinguish and classify bacterial species into two large groups grampositive bacteria and gramnegative bacteria
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There are four basic steps of the Gram stain applying a primary stain (crystal violet) to a heatfixed (death by heat) smear of a bacterial culture;Gramnegative Escherichia coli, the most common Gram stain qualitycontrol bacterium, is decolorized, and is only visible after the addition of the pink counterstain safranin (credit modification of work by Nina Parker)We have cultures of E coli and Bacillus for you to gram stain This will give you gram and gram controls to check your procedure against You can use 2 slides, 1 for each bacterium, or you can divide one slide in half and smear each bacterium on the divided slide
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MORPHOLOGY OF ESCHERICHIA COLI (E COLI) Shape – Escherichia coli is a straight, rod shape (bacillus) bacterium Size – The size of Escherichia coli is about 1–3 µm × 04–07 µm (micrometer) Arrangement Of Cells – Escherichia coli is arranged singly or in pairs Motility – Escherichia coli is a motile bacteriumE coli is described as a Gramnegative bacterium This is because they stain negative using the Gram stain The Gram stain is a differential technique that is commonly used for the purposes of classifying bacteriaE coli S epidermidis Note Escherichia coli is a tiny pink (Gram) rod Staphylococcus epidermidis is a purple (Gram) sphere or coccus Draw a picture of a typical microscopic field and identify both Escherichia coli and Staphylococcus epidermidis Record this in the results section for this lab Colored pencils are available throughout the room
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Thus, for a rodshaped gramnegative bacterium like E coli the comparison with cylindrical soap bubbles has been very helpful To create a cylindrical soap bubble, two fixed rings are needed (Fig1) It is anticipated that the physical properties of the rings and the membrane of the bubble should somehow resemble those of the bacterial polar caps and lateral walls, respectively
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