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Laboratory Exercise 5

Tube Agglutination Test
The Widal Test
Answers to Questions
1. What is the exact meaning of the “titer” of a serum

It is defined as the lowest dilution of the sample that retains a detectable activity (Murray et al.).
The titer of a serum is defined as the highest dilution of serum at which the antigen undergoes
agglutination (Parija, 2012).

2. Differentiate between the following:

Serum: the liquid portion of blood after the clotting factors have been removed and an important
site of antibodies
Antiserum: serum containing antibodies raised against a specific antigen
Antitoxin: protective antibodies that inactivates soluble toxic protein products
(Bauman et al., and Selamawit Debebe,)

3. How would you prepare antiserum for an organism such as E. coli?

E. coli has O, H and K or G antigens; therefore an antiserum should contain antibodies for these
antigens in order to indentify the strains E. coli (Parija, 2012).

4. Indicate the type of antigen (soluble, protein, red blood cells, or bacteria) that is used for
each of the following serological tests:

Agglutination: Specific antibodies or agglutinin bind to surface antigens and caouses visible
clump formation.
Precipitation: Antibody (precipitins) interacts with the soluble antigen in the presence of
electrolyte at a specified pH and temperature to produce a precipitate
Hemolysis: Lysis of RBC observed when using specific antisera
(Parija, 2012)

5. For which one of the above tests is complement necessary?

Hemolysis (Parija, 2012).

Laboratory Exercise 6

amino acids. glucose. amino acids. What is the difference between the components of unheated and heated serum? Unheated serum contains proteins. When heated. 2017). diminished platelet aggregation. impaired macrophage migration. Bactericidal Power of Normal Serum Answers to Questions 1. antibodies. 1978). and lipids. and antibody-dependent. . immunoglobulin aggregation occurs and anti-complementary activities of heat-stable and heat-labile substances are observed. Heat-labile components (such as complement.. 2. and hormones) are destroyed when heated to 56oC 3. What is/are responsible for the bactericidal action of the serum? Heat stable substances of beta-lysin and are independent of complement (C’). growth factors. complement. trypsin inhibitors. hormones. vitamins. and complement fixation which means heated serum has lower bactericidal activity (Soltis et al. Compare the bactericidal activity of heated and unheated serum. This results to inhibition of phagocytosis. and heat- labile substances composed of beta-lysin and are dependent of complement (C’) and antibodies (AB’s) (Tekman. growth factors. cell-mediated cytotoxicity.

. 1996). What are the advantages of RPR over VDRL test? RPR is prepared from a modified VDRL antigen suspensions. 8 Microbiology of the Urinary and Genital Tracts: Urine Culture Techniques . Laboratory Exercise 5 Rapid Plasma Reagin (RPR) Test for Syphilis Answers to Questions 1. and as compared to VDRL test this can be performed in the field... What does RPR determine? It measures IgM and IgG antibodies to lipoidal material released from damaged host cells as well as lipoprotein-like material. 3. 1996). 4.. 2. Therefore. it can test positive. and possibly cardiolipin from the treponemes (Larsen et al. 5. ethyldiaminetetraacetic acid (EDTA) to stabilize the suspension and finely divided charcoal powder as a visualizing agent. These antibodies are produced as a consequence of syphilis and other treponemal diseases. and in response to nontreponemal diseases (Larsen et al. Why are charcoal particles incorporated into the RPR test? They coagglutinate with the antibodies and show up as black clumps against a white card (Larsen et al. What component of the RPR antigen is involved with a reactive patient’s sample? The lipid particles (cardiolipin) of the antigen combine with the antilipoidal antibodies that are present in the sample. but because of nontreponemal diseases (Larsen et al. and it has improved reaction visibility. 1996). 1996). Laboratory Exercise No. it contains ethyl chloride to eliminated the need to heat inactivate serum. Why does a positive RPR not necessarily indicate syphilis infection? The antibodies (antilipoidal antibodies) that are taken into account are produced by not only as a consequence of syphilis but as well as in response to other nontreponemal diseases of an acute and chronic nature in which tissue damage occurs.

This result represents the absence of infection. 3. or presence of blood). How are UTIs acquired/transmitted? Urinary tract infections may be transmitted by improper personal hygiene. If you counted 20 colonies from a 0. Explain why E.. What can you learn from visual inspection of a urine specimen? Visual inspections can be used to diagnose a disease. Cystitis are more often implicated in female because the anus is closer to their urethra. 5.1. or epithelial cells).. and also unusual odor (Brooks et al.01-ml inoculum of a 1:10 dilution of urine. Why is aseptic urine collection important when cultures are ordered? The use of aseptic technique is important to avoid contaminations that may interfere with the accurate identification of bacteria present in the urinary tract (Brooks et al. the number of colonies would be 20. 2013). since the presence of infection is represented by a colony count of 100. bacteria. which means bacteria may be able to get into the bladder more easily (Tortora et al. 2016). however lower colony counts may also dictate infection (AACC. 2013) 2.000 colonies per ml. and infection is associated with a cloudy (presence of other substances such as leukocytes. and contamination during medical procedures. and are often caused by opportunistic pathogens (Tortora et al. colored (due to concentration of solutes... 2016). . 2016). and they have a shorter urethra.000/ml. coli is frequently implicated in cystitis in females. how many organisms per milliliter of specimen would you report? Is this number significant? Represented in milliliter. 4.

while yeasts have a lengthier generation time (Kumar.. 9 Culture and Microscopic Examination of Fungi 1. What are some similarities and differences of the yeasts that were cultured from your mouth? N/A 4. Can bacteriological media be used for the cultivation of molds? Explain your answer. 5. and budding is an asexual reproduction that begins as a protuberance from the parent cell and grows to become a daughter cell (Tortora et al. . so bacteriological media that contain glucose and modified peptone with a neutral pH may also be used to cultivate molds. Define budding and hyphae. 3. Bacteria reproduce rapidly through binary fission. Hyphae is a long filament of fungal cells or actinomycetes. 2016) 2. while aerial hyphae project the surface of the mycelium and bear the reproductive structures of the mold which produces asexual pores (Kumar. Laboratory Exercise No. 2012). Why do yeasts generally have to be cultured for longer periods than most bacteria? Fungi are eukaryotic protists which are more complex and differ in many ways with bacteria such as in the time it needs to culture fungi. 2012). What is the difference between vegetative and aerial mycelia? A mycelium is constituted by a tangled mass of hyphae that forms in molds or filamentous fungi. A vegetative mycelium is a hypha that penetrates the supporting medium and absorb nutrients. Yes. Most fungi occur in nature and grow readily on simple sources of nitrogen and carbohydrate.

2. . Laboratory Exercise No. gamma-rays or UV light (Tortora et al. What is a Mutagen? Give several examples. benzopyrene and aflatoxin. It requires the binding of streptomycin to its protein to function properly during protein synthesis (Gregory et al. 2012).. aureus. 3. radiations may be X-rays. Mutagens are environmental agents such as chemicals or radiation. 2016). How would you describe a streptomycin-dependent mutant bacterium? It is a mutant bacterium that harbors resistance to streptomycin which is due to the substitution of a single amino acid (rpsL gene encoding the S12 ribosomal protein). 4. 2001). Antimicrobial use and misuse. 10 Bacterial Mutation 1.. How can bacterial mutations occur? This may result from the exposure of the bacterium to radiation or chemicals (antibiotics) that cause changes in the bacterial DNA (Kumar. nucleoside analogue. and factors that enhance its transmission may be increase the frequency of mutant S. Chemical mutagens include nitrous acid. What methods could be used to increase the frequency of the appearance of mutant Staphylococcus aureus.