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Exam Number : EX200
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EX200 Exam Format | EX200 Course Contents | EX200 Course Outline | EX200 Exam Syllabus | EX200 Exam Objectives

Understand and use essential tools

Access a shell prompt and issue commands with correct syntax

Use input-output redirection (>, >>, |, 2>, etc.)

Use grep and regular expressions to analyze text

Access remote systems using SSH

Log in and switch users in multiuser targets

Archive, compress, unpack, and uncompress files using tar, star, gzip, and bzip2

Create and edit text files

Create, delete, copy, and move files and directories

Create hard and soft links

List, set, and change standard ugo/rwx permissions

Locate, read, and use system documentation including man, info, and files in /usr/share/doc

Operate running systems

Boot, reboot, and shut down a system normally

Boot systems into different targets manually

Interrupt the boot process in order to gain access to a system

Identify CPU/memory intensive processes and kill processes

Adjust process scheduling

Manage tuning profiles

Locate and interpret system log files and journals

Preserve system journals

Start, stop, and check the status of network services

Securely transfer files between systems

Configure local storage

List, create, delete partitions on MBR and GPT disks

Create and remove physical volumes

Assign physical volumes to volume groups

Create and delete logical volumes

Configure systems to mount file systems at boot by universally unique ID (UUID) or label

Add new partitions and logical volumes, and swap to a system non-destructively

Create and configure file systems

Create, mount, unmount, and use vfat, ext4, and xfs file systems

Mount and unmount network file systems using NFS

Extend existing logical volumes

Create and configure set-GID directories for collaboration

Configure disk compression

Manage layered storage

Diagnose and correct file permission problems

Deploy, configure, and maintain systems

Schedule tasks using at and cron

Start and stop services and configure services to start automatically at boot

Configure systems to boot into a specific target automatically

Configure time service clients

Install and update software packages from Red Hat Network, a remote repository, or from the local file system

Work with package module streams

Modify the system bootloader

Manage basic networking

Configure IPv4 and IPv6 addresses

Configure hostname resolution

Configure network services to start automatically at boot

Restrict network access using firewall-cmd/firewall

Manage users and groups

Create, delete, and modify local user accounts

Change passwords and adjust password aging for local user accounts

Create, delete, and modify local groups and group memberships

Configure superuser access

Manage security

Configure firewall settings using firewall-cmd/firewalld

Create and use file access control lists

Configure key-based authentication for SSH

Set enforcing and permissive modes for SELinux

List and identify SELinux file and process context

Restore default file contexts

Use boolean settings to modify system SELinux settings

Diagnose and address routine SELinux policy violations

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abstract of Biochemical exams | Resources

assessments used to determine Gram tremendous micro organism checks used to establish Gram negative bacteria Mannitol Salt Agar (MSA)

This category of medium is each selective and differential. The MSA will opt for for organisms similar to Staphylococcus species which may are living in areas of excessive salt concentration (plate on the left within the photograph below). here's in distinction to Streptococcus species, whose growth is selected against by this excessive salt agar (plate on the correct in the image beneath).

The differential ingredient in MSA is the sugar mannitol. Organisms able to using mannitol as a meals supply will produce acidic byproducts of fermentation with a view to lessen the pH of the media. The acidity of the media will trigger the pH indicator, phenol crimson, to show yellow. Staphylococcus aureus is able to fermenting mannitol (left facet of left plate) while Staphylococcus epidermidis isn't (appropriate aspect of left plate).



Glucose broth with Durham tubes

this is a differential medium. It exams an organism's capability to ferment the sugar glucose in addition to its means to transform the conclusion manufactured from glycolysis, pyruvic acid into gaseous byproducts. here's a look at various usual when trying to establish Gram-poor enteric micro organism, all of which are glucose fermenters but handiest a few of which produce fuel.

Like MSA, this medium also carries the pH indicator, phenol red. If an organism is able to fermenting the sugar glucose, then acidic byproducts are fashioned and the pH indicator turns yellow. Escherichia coli is in a position to fermenting glucose as are Proteus mirabilis (some distance right) and Shigella dysenteriae (far left).  Pseudomonas aeruginosa (center) is a nonfermenter.

The end made from glycolysis is pyruvate. Organisms that are capable of changing pyruvate to formic acid and formic acid to H2 (g) and CO2 (g), by means of the motion of the enzyme formic hydrogen lyase, emit gas. This gasoline is trapped within the Durham tube and appears as a bubble on the correct of the tube. Escherichia coli and Proteus mirabilis (some distance right) are both gasoline producers. word that Shigella dysenteriae (a long way left) ferments glucose but doesn't produce gas.

*be aware - broth tubes may also be made containing sugars apart from glucose (e.g. lactose and mannitol).  because the identical pH indicator (phenol crimson) is also used in these fermentation tubes, the identical outcomes are considered wonderful  (e.g. a lactose broth tube that turns yellow after incubation has been inoculated with an organism that can ferment lactose).


Blood Agar Plates (BAP)

this is a differential medium. it's a wealthy, advanced medium that consists of 5% sheep pink blood cells. BAP checks the ability of an organism to supply hemolysins, enzymes that hurt/lyse red blood cells (erythrocytes). The diploma of hemolysis by way of these hemolysins is useful in differentiating contributors of the genera Staphylococcus, Streptococcus and Enterococcus.

  • Beta-hemolysis is finished hemolysis. it's characterised through a clear (transparent) zone surrounding the colonies. Staphylococcus aureus, Streptococcus pyogenes and Streptococcus agalactiae are b-hemolytic (the photo on the left below indicates the beta-hemolysis of S. pyogenes).
  • Partial hemolysis is termed alpha-hemolysis. Colonies typically are surrounded via a green, opaque zone. Streptococcus pneumoniae and Streptococcus mitis are a-hemolytic (the graphic on the appropriate under suggests the a-hemolysis of S. mitis).
  • If no hemolysis happens, here's termed gamma-hemolysis. There aren't any amazing zones across the colonies. Staphylococcus epidermidis is gamma-hemolytic.
  • Picture of Beta HemolysisPicture of Alpha Hemolysis

    What class of hemolysis is seen on each and every considered one of right here plates?


    Streak-stab method

    frequently when inoculating a BAP to have a look at hemoloysis patterns, investigators will also stab several instances throughout the agar using an inoculating loop. This stab makes it possible for for the detection of streptolysin O, a selected hemolysin produced with the aid of Streptococcus pyogenes. This hemolysin is inactivated by means of O2 and is just seen subsurface (in an anaerobic atmosphere) across the stab mark. be aware the oval-formed areas of clearing across the stab marks in the image below; these are led to by way of streptolysin O.

    Bile Esculin Agar

    this is a medium it's each selective and differential. It exams the skill of organisms to hydrolyze esculin in the presence of bile. it is commonly used to determine participants of the genus Enterococcus (E faecalis and E. faecium).

    the primary selective ingredient in this agar is bile, which inhibits the growth of Gram-positives other than enterococci and a few streptococci species. The 2nd selective ingredient is sodium azide. This chemical inhibits the increase of Gram-negatives.

    The differential ingredient is esculin. If an organism can hydrolyze esculin within the presence of bile, the product esculetin is shaped. Esculetin reacts with ferric citrate (within the medium), forming a phenolic iron advanced which turns the total slant darkish brown to black. The tube on the a ways right was inoculated with E. faecalis (advantageous). The tube within the middle was inoculated with a bilie esculin terrible organism and the tube on the left was uninoculated.

    Bile Esculin Agar


    Sulfur Indole Motility Media (SIM)

    this is a differential medium. It checks the capability of an organism to do a number of issues: reduce sulfur, produce indole and swim through the agar (be motile). SIM is universal to distinguish participants of Enterobacteriaceae.

    Sulfur will also be reduced to H2S (hydrogen sulfide) either by using catabolism of the amino acid cysteine by the enzyme cysteine desulfurase or via discount of thiosulfate in anaerobic respiratory. If hydrogen sulfide is produced, a black colour varieties in the medium. Proteus mirabilis is effective for H2S production. The organism pictured on the some distance left is advantageous for hydrogen sulfide construction.

    micro organism that have the enzyme tryptophanase, can convert the amino acid, tryptophane to indole. Indole reacts with brought Kovac’s reagent to form rosindole dye which is purple in color (indole +). Escherichia coli is indole high quality. The organism pictured 2nd from left is E. coli and is indole positive.

    SIM tubes are inoculated with a single stab to the backside of the tube. If an organism is motile than the growth will radiate from the stab mark and make the whole tube seem turbid. Pseudomonas aeruginosa and the pressure of Proteus mirabilis that we work with are motile.

    Sulfur Indole Motility Media


    Kliger’s Iron Agar (KIA)

    this is a differential medium. It tests for organisms’ expertise to ferment glucose and lactose to acid and acid plus fuel end items. It additionally enables for identification of sulfur reducers. This media is wide-spread to separate lactose fermenting contributors of the family Enterobacteriaceae (e.g. Escherichia coli) from contributors that do not ferment lactose, like Shigella dysenteriae. These lactose nonfermenting enterics often are usually the greater severe pathogens of the the gastrointestinal tract.

    the primary differential ingredient, glucose, is in very short supply. Organisms able to fermenting this sugar will use it up in the first few hours of incubation. Glucose fermentation will create acidic byproducts that allows you to turn the phenol pink indicator within the media yelllow. consequently, after the primary few hours of incubation, the tube should be entirely yellow. At this point, when the glucose has been all used up, the organism should opt for another food supply. If the organism can ferment lactose, this is the sugar it's going to select. Lactose fermentation will continue to provide acidic byproducts and the media will continue to be yellow (photograph on the a long way left beneath). If gasoline is produced as a result of glucose or lactose fermentation, then fissures will appear within the agar or the agar will be lifted off the backside of the tube.

    If an organism cannot use lactose as a food supply it may be pressured to make use of the amino acids / proteins in the media. The deamination of the amino acids creates NH3, a vulnerable base, which motives the medium to turn into alkaline. The alkaline pH explanations the phenol pink indicator to start to show red. for the reason that the incubation time is brief (18-24 h), simplest the slant has an opportunity to show red and never the complete tube. consequently an organism that can ferment glucose however now not lactose, will produce a crimson slant and a yellow butt in a KIA tube (2nd from the left beneath). These organisms are the more severe pathogens of the GIT similar to Shigella dysenteriae.

    If an organism is capable of using neither glucose nor lactose, the organism will use totally amino acids / proteins. The slant of the tube may be purple and the color of the butt will stay unchanged (graphic on the far correct under). Pseudomonas aeruginosa is an illustration of a nonfermenter.

    KIA tubes are also capable of detecting the construction of H2S. it's considered as a black precipitate (second photograph from the right). now and again the black precipitate obscures the butt of the tube. In such instances, the organisms should be regarded high-quality for glucose fermentation (yellow butt). Proteus mirabilis (pictured right here, second from appropriate) is a glucose superb, lactose poor, sulfur reducing enteric.


    Nitrate Broth

    this is a differential medium. it's used to check if an organism is able to cutting back nitrate (NO3-) to nitrite (NO2-) or other nitrogenous compounds by the use of the action of the enzyme nitratase (also known as nitrate reductase). This look at various is important within the identification of both Gram-nice and Gram-negative species.After incubation, these tubes are first inspected for the presence of gasoline within the Durham tube. in the case of nonfermenters, this is indicative of discount of nitrate to nitrogen fuel. however, in many circumstances fuel is produced via fermentation and extra trying out is integral to examine if reduction of nitrate has occurred. This further trying out comprises the addition of sulfanilic acid (commonly called nitrate I) and dimethyl-alpha-napthalamine (nitrate II). If nitrite is existing in the media, then it will react with nitrate I and nitrate II to kind a crimson compound. this is regarded a favorable outcomes. If no crimson colour varieties upon addition of nitrate I and II, this indicates that either the NO3- has no longer been transformed to NO2- (a negative effect), or that NO3- became converted to NO2- and then instantly decreased to some other, undetectable type of nitrogen (also a good outcomes). in an effort to assess which of the previous is the case, elemental zinc is brought to the broth. Zinc will convert any remaining NO3- to NO2- thus permitting nitrate I and nitrate II to react with the NO2- and kind the purple pigment (a tested bad outcome). If no color exchange happens upon addition of zinc then this capacity that the NO3- become transformed to NO2- after which become transformed to every other undetectable form of nitrogen (a favorable effect).

    If the nitrate broth turns purple (tubes pictured in the core) after nitrate I and nitrate II are delivered, this color indicates a favorable influence. If instead, the tube turns purple (tube pictured on the left) after the addition of Zn, this indicates a negative outcome. If there is no colour change within the tube after the addition of nitrate I and nitrate II, the influence is uncertain. If the tube is colorless (graphic on the right) after the addition of Zn this suggests a positive verify.

    Nitrate Broth


    Catalase look at various

    This look at various is used to establish organisms that produce the enzyme, catalase. This enzyme detoxifies hydrogen peroxide via breaking it down into water and oxygen gasoline.

    The bubbles on account of production of oxygen gasoline evidently point out a catalase high-quality outcomes. The sample on the right below is catalase tremendous. The Staphylococcus spp. and the Micrococcus spp. are catalase fantastic. The Streptococcus and Enterococcus spp. are catalase poor.


    Oxidase examine

    This check is used to determine microorganisms containing the enzyme cytochrome oxidase (important in the electron transport chain). it's normal to differentiate between oxidase bad Enterobacteriaceae and oxidase positive Pseudomadaceae.

    Cytochrome oxidase transfers electrons from the electron transport chain to oxygen (the final electron acceptor) and reduces it to water. in the oxidase check, synthetic electron donors and acceptors are provided. When the electron donor is oxidized via cytochrome oxidase it turns a gloomy pink. here is considered a good effect. within the graphic below the organism on the correct (Pseudomonas aeruginosa) is oxidase fantastic.



    Coagulase look at various

    Coagulase is an enzyme that clots blood plasma. This examine is performed on Gram-superb, catalase fine species to establish the coagulase superb Staphylococcus aureus. Coagulase is a virulence element of S. aureus. The formation of clot round an infection brought about with the aid of this bacteria seemingly protects it from phagocytosis. This examine differentiates Staphylococcus aureus from different coagulase terrible Staphylococcus species.

    coagulase tubesstaph aureus


    Taxos A (bacitracin sensitivity testing)

    here is a differential look at various used to differentiate between organisms delicate to the antibiotic bacitracin and people now not. Bacitracin is a peptide antibiotic produced via Bacillus subtilis. It inhibits phone wall synthesis and disrupts the cellphone membrane. This look at various is commonly used to distinguish between the b-hemolytic streptococci: Streptococcus agalactiae (bacitracin resistant) and Streptococcus pyogenes (bacitracin sensitive). The plate under become streaked with Streptococcus pyogenes; note the big zone of inhibition surrounding the disk.



    Taxos P (optochin sensitivity testing)

    this is a differential test used to distinguish between organisms sensitive to the antibiotic optochin and people not. This check is used to differentiate Streptococcus pneumoniae (optochin sensitive (pictured on the appropriate beneath)) from different a-hemolytic streptococci (optochin resistant (Streptococcus mitis is pictured on the left beneath)).


    MacConkey agar

    This medium is each selective and differential. The selective materials are the bile salts and the dye, crystal violet which inhibit the increase of Gram-high quality bacteria. The differential ingredient is lactose. Fermentation of this sugar outcomes in an acidic pH and explanations the pH indicator, neutral crimson, to show a bright pinky-pink color. as a result organisms capable of lactose fermentation equivalent to Escherichia coli, form shiny pinky-crimson colonies (plate pictured on the left right here). MacConkey agar is prevalent to distinguish between the Enterobacteriaceae.

    MacConkey PositiveMacConkey NegativeOrganism on left is wonderful for lactose fermentation and that on the appropriate is bad.


    Simmon’s Citrate Agar

    here is a defined medium used to verify if an organism can use citrate as its sole carbon supply. it is often used to distinguish between members of Enterobacteriaceae. In organisms able to employing citrate as a carbon source, the enzyme citrase hydrolyzes citrate into oxaoloacetic acid and acetic acid. The oxaloacetic acid is then hydrolyzed into pyruvic acid and CO2. If CO2 is produced, it reacts with add-ons of the medium to produce an alkaline compound (e.g. Na2CO3). The alkaline pH turns the pH indicator (bromthymol blue) from green to blue. here's a good result (the tube on the right is citrate high-quality). Klebsiella pneumoniae and Proteus mirabilis are examples of citrate superb organisms. Escherichia coli and Shigella dysenteriae are citrate bad.

    Simmon's Citrate Agar


    Spirit Blue agar

    This agar is used to identify organisms that are in a position to producing the enzyme lipase. This enzyme is secreted and hydrolyzes triglycerides to glycerol and three long chain fatty acids. These compounds are small enough to circulate during the bacterial mobilephone wall. Glycerol will also be converted right into a glycolysis intermediate. The fatty acids may also be catabolized and their fragments can ultimately enter the Kreb’s cycle. Spirit blue agar incorporates an emulsion of olive oil and spirit blue dye. bacteria that produce lipase will hydrolyze the olive oil and produce a halo around the bacterial boom. The Gram-wonderful rod, Bacillus subtilis is lipase wonderful (pictured on the appropriate) The plate pictured on the left is lipase terrible.

    Spirit Blue


    Starch hydrolysis verify

    This examine is used to determine bacteria that can hydrolyze starch (amylose and amylopectin) the usage of the enzymes a-amylase and oligo-1,6-glucosidase. frequently used to differentiate species from the genera Clostridium and Bacillus. because of the enormous measurement of amylose and amylopectin molecules, these organisms cannot pass in the course of the bacterial telephone wall. so as to use these starches as a carbon supply, micro organism need to secrete a-amylase and oligo-1,6-glucosidase into the extracellular space. These enzymes wreck the starch molecules into smaller glucose subunits which may then enter at once into the glycolytic pathway. with a purpose to interpret the effects of the starch hydrolysis verify, iodine ought to be delivered to the agar. The iodine reacts with the starch to kind a gloomy brown colour. thus, hydrolysis of the starch will create a transparent zone across the bacterial increase. Bacillus subtilis is wonderful for starch hydrolysis (pictured under on the left). The organism proven on the correct is poor for starch hydrolysis.


    Methyl pink / Voges-Proskauer (MR/VP)

    This look at various is used to verify which fermentation pathway is used to make the most of glucose. within the blended acid fermentation pathway, glucose is fermented and produces a couple of biological acids (lactic, acetic, succinic, and formic acids). The solid construction of adequate acid to beat the phosphate buffer will outcomes in a pH of under four.four. If the pH indicator (methyl red) is introduced to an aliquot of the culture broth and the pH is beneath 4.four, a purple colour will appear (first photograph, tube on the left). If the MR turns yellow, the pH is above 6.0 and the blended acid fermentation pathway has not been utilized (first photograph, tube on the right). the two,three butanediol fermentation pathway will ferment glucose and produce a 2,three butanediol conclusion product as an alternative of biological acids. with the intention to look at various this pathway, an aliquot of the MR/VP culture is eliminated and a-naphthol and KOH are added. they are shaken together vigorously and set aside for approximately one hour except the results can also be read. The Voges-Proskauer examine detects the presence of acetoin, a precursor of two,three butanediol. If the culture is nice for acetoin, it's going to turn “brownish-red to pink” (tube on the left in the 2nd picture). If the subculture is bad for acetoin, it's going to flip “brownish-green to yellow” (tube on the left within the 2nd graphic). word: A tradition will always handiest be fine for one pathway: either MR+ or VP+. Escherichia coli is MR+ and VP-. In distinction, Enterobacter aerogenes and Klebsiella pneumoniae are MR- and VP+. Pseudomonas aeruginosa is a glucose nonfermenter and is consequently MR- and VP-.



    CAMP test

    CAMP element is a diffusible, warmth-stable protein produced with the aid of community B streptococci. here's a synergistic test between Staphylococcus aureus and Streptococcus agalactiae. S. agalactiae produces CAMP ingredient. S. aureus produces sphingomyelin C, which binds to purple blood telephone membranes. both micro organism are streaked at 90o angles of 1 an extra. They do not contact. The CAMP component produced via S. agalactiae enhances the beta-hemolysis of S. aureus by way of binding to already broken red blood cells. in consequence, an arrow of beta-hemolysis is produced between the two streaks. The examine is presumptive for S. agalactiae that produces CAMP factor.

    in the image here, Streptococcus agalactiae became streaked all over the right region of the plate and brought down toward the center of the plate. Staphylococcus aureus turned into streaked in a straight line throughout the core of the plate. Rings of hemolysis are evident throughout S. aureus, despite the fact the hemolysis if greatly more suitable (in an arrow form) where the S. agalactiae crosses the hemolysis rings.

    CAMP Test


    Urease test

    This look at various is used to identify bacteria capable of hydrolyzing urea using the enzyme urease. it is commonly used to differentiate the genus Proteus from other enteric bacteria. The hydrolysis of urea types the vulnerable base, ammonia, as one among its products. This susceptible base raises the pH of the media above 8.four and the pH indicator, phenol red, turns from yellow to crimson. Proteus mirabilis is a speedy hydrolyzer of urea (core tube pictured right here). The tube on the a long way appropriate became inoculated with a urease poor organism and the tube on the far left become uninoculated.

    Urease Test


    Motility agar

    is a differential medium used to investigate whether an organism is fitted with flagella and accordingly able to swimming far from a stab mark. The consequences of motility agar are sometimes problematic to interpret. frequently, if the total tube is turbid, this shows that the micro organism have moved far from the stab mark (are motile). The organisms in the two tubes pictured on the appropriate are motile. If, despite the fact, the stab mark is certainly visible and the relaxation of the tube is not turbid, the organism is probably going nonmotile (tube pictured on the left).

    Motility Agar


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