Staphylococcus saprophyticus biochemical test results

Summary of Biochemical Tests

Staphylococcus saprophyticus is one of the most frequently encountered microorganisms associated with acute urinary tract infections UTIs in young, sexually active female outpatients. Conventional identification methods based on biochemical characteristics can efficiently identify S. Rapid and direct identification of this bacterium from urine samples would be useful to improve time required for the diagnosis of S.

We have developed a PCR-based assay for the specific detection of S. An arbitrarily primed PCR amplification product of bp specific for S. The PCR assay was specific for S. This assay was also able to amplify efficiently DNA from all 60 strains of S. This assay was adapted for direct detection from urine samples.

The sensitivity levels achieved with urine samples was 19 CFU with 30 cycles of amplification and 0. This PCR assay for the specific detection of S. Coagulase-negative staphylococci are commensal organisms of human skin flora but have become major etiological agents of nosocomial bacteremia and can colonize a variety of medical devices However, Staphylococcus saprophyticus is the second most frequently encountered agent of acute urinary tract infections UTIs after Escherichia coli 10 These coagulase-negative staphylococci are rarely found as a cause of UTIs in hospitalized patients or as a contaminant of urine cultures 15 and are characterized by the low bacterial counts less than 10 5 CFU per ml required to elicit a UTI The use of spermicide-coated condoms has now been associated with an increased risk of UTIs caused by S.

However, these systems require at least 20 h for staphylococcal species identification and occasionally misidentify S. However, these assays have not been evaluated for direct detection of S.

Although S. A rapid and sensitive DNA-based assay which is specific for S. In this study, we present the development of an S. This S. This assay will be combined in multiplex with other PCR assays currently under development in our laboratory to allow the concomitant detection of other bacteria frequently associated with UTIs. Three S. The strains were cultured on sheep blood agar or in brain heart infusion BHI medium. Forty-nine gram-positive, 31 gram-negative, and 60 clinical isolates were used to establish the performance of the S.

A total of five culture-negative urine specimens received at the Microbiology Laboratory of CHUL, all collected from different patients, were used in this study.This type of medium is both selective and differential.

The MSA will select for organisms such as Staphylococcus species which can live in areas of high salt concentration plate on the left in the picture below. This is in contrast to Streptococcus species, whose growth is selected against by this high salt agar plate on the right in the picture below.

The differential ingredient in MSA is the sugar mannitol. Organisms capable of using mannitol as a food source will produce acidic byproducts of fermentation that will lower the pH of the media.

The acidity of the media will cause the pH indicator, phenol red, to turn yellow. Staphylococcus aureus is capable of fermenting mannitol left side of left plate while Staphylococcus epidermidis is not right side of left plate. This is a differential medium. It tests an organism's ability to ferment the sugar glucose as well as its ability to convert the end product of glycolysis, pyruvic acid into gaseous byproducts.

staphylococcus saprophyticus biochemical test results

This is a test commonly used when trying to identify Gram-negative enteric bacteria, all of which are glucose fermenters but only some of which produce gas.

Like MSA, this medium also contains the pH indicator, phenol red. If an organism is capable of fermenting the sugar glucose, then acidic byproducts are formed and the pH indicator turns yellow. Escherichia coli is capable of fermenting glucose as are Proteus mirabilis far right and Shigella dysenteriae far left. Pseudomonas aeruginosa center is a nonfermenter. The end product of glycolysis is pyruvate. Organisms that are capable of converting pyruvate to formic acid and formic acid to H 2 g and CO 2 gvia the action of the enzyme formic hydrogen lyase, emit gas.

This gas is trapped in the Durham tube and appears as a bubble at the top of the tube. Escherichia coli and Proteus mirabilis far right are both gas producers. Notice that Shigella dysenteriae far left ferments glucose but does not produce gas.

The degree of hemolysis by these hemolysins is helpful in differentiating members of the genera StaphylococcusStreptococcus and Enterococcus.

staphylococcus saprophyticus biochemical test results

Often when inoculating a BAP to observe hemoloysis patterns, investigators will also stab several times through the agar using an inoculating loop. This stab allows for the detection of streptolysin O, a specific hemolysin produced by Streptococcus pyogenes.

This hemolysin is inactivated by O 2 and is only seen subsurface in an anaerobic environment around the stab mark.A Microbial Biorealm page on the genus Staphylococcus saprophyticus. NCBI: Taxonomy. Staphylococcus saprophyticus is a Gram-positive, coagulase-negative facultative species of Staphylococcus, which is a leading cause of cystisis in women and is associated with uncomplicated urinary tract infection UTI in humans.

Like other Staphylococci, S. Young women are more susceptible to colonization in the urinary tracts and sexual intercourse promotes its spread. This antigen was later classified as S. As ofthe genome of S. The genome was sequenced to better understand the pathenogenesis of the organism.

Whole-genome shotgun sequencing was used, sequencing in kb or 10 kb inserts. The gaps were filled by PCR direct sequencing, using specific primers at the ends of each gap. The genome of S. The SCCs are thought to have integrated into the genome through a two-step process and contain a restriction enzyme modification system and a cassette chromosome recombinase Ccr.

The genomic island in S. The two plasmids carry the gene for an aquaporin which regulates the osmolarity of the cell.

Multiple copies of the plasmids provide for expression of more water channels. Staphylococcus saprophyticus is a coagulase-negative species of Staphylococcus.

Like other Staphylococci, it is Gram-positive, is globular shaped, and is a facultative anaerobe. It has abundant transporter systems to adapt to ever changing pH, osmolarity, and concentration of urea in human urine. The plasmids contain a gene coding for aquaporin Z. The amount of water channels created is regulated by the number of copies of the plasmids.

It has both a pH-driven symporter and a sodium-dependent symporter to transport divalent cations, including iron, into the cell. This is how the cell metabolizes Nitrogen.

Biochemical Tests for Streptococcus Organisms

Urease activity is known to be an infection causing factor in UTIs. The antibody to the poplypeptide inhibits hemagglutination. Staphylococcus saprophyticus adheres to uroepithelial cells and sheep erythrocytes causing hemoglutination. Spermicides and candidal infections affect the vaginal flora, increasing the risk of infection.

Staphylcoccus saprophyticus is not naturally found in healthy humans. It infects humans through sexual intercourse or through contact with animals.Streptococcus organisms are Gram positive, microaerophilic and non-motile bacteria. There exist several sources of streptococcus organisms that include humans as well as many animals where they mostly form colonies in the mucosal surface of the mouth, pharynx, etc.

When they are found in drinking water then the water is aid to be contaminated with fecal matters. Some of the food materials that may be infected with streptococcus are milk, egg, ground ham etc. Some of the foods which are also left for a long time at room temperature before eating are also infected with streptococcus organisms.

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So, the basic objective of this test is to study about various biochemical tests for Streptococcus organisms. MR-VP Broth. Nutrient gelatin medium.

Novobiocin Susceptibility Test: Principle, procedure and interpretations

Simmon citrate agar. Urea Agar. Alpha naphthylamine reagent. Zinc dust. Baritt reagent A — 5G Alpha naphthol in ml absolute ethyl alcohol. Baritt reagent B — Potassium Hydroxide gm in ml Distilled water.

Methyl red reagent.

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Miscellaneous: Inoculating loop. Test tubes. Then incubate the plate at 35oC for about 24 hours. If a clear zone is formed around the colony then it beta-hemolytic.

A streptococcus organism is beta-hemolytic. Catalase test. With the help of inoculating tube, take a little amount of 24 hours culture of test organism and dip it in the test tube. Then keep this test tube in a dark background and see if there is any bubble formation in the tip of inoculation tube.

If bubble is formed then it is catalase positive and if no bubble formation then it is catalase negative. Staphylococcus gives negative test for Catalase test. Gelatin hydrolysis. Take nutrient gelatin medium in a tube and to it add the test bacteria to it by stabbing. In another tube take nutrient gelatin medium and leave it uninoculated for standard.

Up to then everyday bring out the tubes and keep it in ice bath or refrigerator for about min for checking gelatin liquefaction. The purpose of keeping it in refrigerator is because in general gelatin liquefies at 28 o C, therefore to confirm that it is liquefying because of the bacteria it is kept at a low temperature in a refrigerator.

You can confirm hydrolyzation by tilting the tubes a bit.

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Staphylococcus is gelatin positive. H 2 S production. Take a labelled tube containing SIM medium and inoculate the organism by stabbing.We studied the biochemical properties of the urease of Staphylococcus saprophyticus and the possible role of the urease in experimental urinary tract infections.

For this purpose, the nonhemagglutinating and nonadherent strainwhich was isolated from a case of symptomatic urinary tract infection, was used. The urease was shown to have a Km of 6. The enzyme was inhibited by acetohydroxamic acid in a noncompetitive manner.

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To assess the contribution of S. In the rat model of ascending unobstructed urinary tract infection, higher numbers of CFU. Moreover, bladder stones were found in animals infected with the urease-positive strain only. Interestingly, the difference in mean bacterial counts of the bladders was found to be significant by the Wilcoxon two-sample test P less than 0.

Immunoblot studies revealed a faint antibody response in rats infected with the mutant strain, although bacteria could still be detected in the kidneys after 7 days. Sera of animals challenged with the parent strain reacted strongly with many antigens of S. Our data indicate that urease is a major factor for invasiveness of S.

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Abstract We studied the biochemical properties of the urease of Staphylococcus saprophyticus and the possible role of the urease in experimental urinary tract infections. Images in this article Image on p. Image on p. Comparison of adherence and urine growth rate properties of Staphylococcus saprophyticus and Staphylococcus epidermidis.

Eur J Clin Microbiol. A rapid, sensitive method for detection of alkaline phosphatase-conjugated anti-antibody on Western blots. Anal Biochem. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Role of bacterial urease in experimental pyelonephritis. J Bacteriol. Surface properties of Staphylococcus saprophyticus and Staphylococcus epidermidis as studied by adherence tests and two-polymer, aqueous phase systems.

Acta Pathol Microbiol Scand B. Jack bean urease EC 3. The molecular size and the mechanism of inhibition by hydroxamic acids. Spectrophotometric titration of enzymes with reversible inhibitors.Staphylococcus Saprophyticus Biochemical Test Results. Staphylococcus epidermidis may cause infection of conjunctiva conjunctivitiscornea keratitis or hair follicles on the edge of the eyelid folliculitis, stye 1. I am trying to identify an unknown bacteria for my micro lab.

May-Jun; 6 3 : Highly suspected microbes: Escherichia coli and Staphylococcus saprophyticus. Name: Dosage: Linezolid: - mgs: twice a day for Most non-pathogenic staphylococci will not ferment mannitol. Reference number It produces toxic shock syndrome toxin, exfoliative toxin, and enterotoxin. Learn vocabulary, terms, and more with flashcards, games, and other study tools. Most of them are harmless and reside normally in the skin and mucous membrane of human beings along with other organisms.

However, the efficacy of nitrofurantoin in treating clinical. A tech is reading the biochemical tests for identifying a particular gram-negative rod. The coagulase test can be performed using two different procedures - Slide test and tube test. Biochemical Test.

All the 18 Staphylococcus saprophyticus strains were identified by their colony morphology, biochemical tests and were subjected to antibiotic sensitivity testing by modified Kirby-bauer disc diffusion method. Consult appropriate references for additional differential tests.

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Of the 14 sam-ples of Staphylococcus aureus, there were eight samples of cefoxitin-resistant samples. Correlation of culture with urinalysis results J. Additionally, the urease reactivity of sera from patients diagnosed with brucellosis was.

It is possible for gram-positive bacteria to grow, but it would barely grow. Prior to this, the presence of coagulase-negative staphylococci CoNS in urine specimens was dismissed as contamination. The sequence and phylogenetic analysis of PCR amplified. Biochemical tests indicate this microorganism also carries out a weakly positive reaction to the nitrate reductase test. Staphylococcus saccharolyticus is a Gram-positive, coagulase-negative, anaerobic member of the bacterial genus Staphylococcus consisting of single and clustered cocci.

After these biochemical test were performed and recorded from the identification chart that was handed out by the professor the result of Unknown A is Staphylococcus epidermidis.

Staphylococcus saprophyticus is considered Normal Flora.

staphylococcus saprophyticus biochemical test results

The key difference between epidermidis and aureus is that Staphylococcus epidermidis is a non-haemolytic bacterium while Staphylococcus aureus is a haemolytic bacterium. These tests identify S.It is usually performed on urine isolates isolated from women of reproductive age groups that are coagulase negative. So when coagulase negative staphylococcus is isolated from a young sexually active womenlaboratory must further identify that isolate and find out is this a true pathogen Staphylococcus saprophyticus-novobiocin resistant or a contaminant i.

Staphylococcus epidermidis-novobiocin sensitive. Therefore, screening coagulase-negative staphylococci from urine cultures for novobiocin resistance is a reliable presumptive identification of S. Novobiocin susceptibility test can be done using a tube method and it takes around 5 hours to get the results. Testing should be performed on isolated colonies of aerobic, catalase -positive, coagulase -negative gram positive cocci.

Novobiocin sensitivity testing A. Novobiocin-resistant S. Novobiocin Sensitive S. The novobiocin disk is not helpful and can give misleading results if it is performed on isolates other that those from urinary specimens. Occasional human isolates that are not S. Salt Tolerance Test is used in the identification of enterococcal group D Streptococcus on the basis of their salt tolerance.

The ability of the bacteria to grow in the presence of variable amount of Sodium […]. Pyrrolidonyl Arylamidase PYR test is a rapid test which is used for the presumptive identification of group A beta-hemolytic Streptococci and Enterococci.

Staphylococcus Saprophyticus Biochemical Test Results

PYR test is available in different format. Original PYR test used to take […]. Coagulase is an enzyme produced by S. Staphylococcus aureus produces two […]. Thank you for your concern and comment. We prepare the agar plates using commercially available dehydrated media and we purchase Novobiocin disc from the suppliers.

Anonymous Ji As novobiocin is not included in the CLSI chart, there is slight variation regarding zone size interpretation criteria among manufacture of the disk. As this drug is not used for the therapeutic purpose but only for the identification of species of the genera Staphylococcus designating intermediate to an isolate makes no sense.

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Remember, While performing the test, please follow zone size interpretative chart provided by the manufacture of the disk. I complete an experiment but was not sure of my results. Is it effective against S. How about Penicillin and gentamicin are they resistant?

staphylococcus saprophyticus biochemical test results

Pls explain. I appreciate your help.