| What alternative method is suggested if incineration is not possible? | not possible deep burial may be a viable option | Ensure that an adequate amount of fuel is added to completely reduce the carcass to ash. If incineration is not possible deep burial may be a viable option. Incase of prolonged rains or logistical problem, carcasses disposal may be delayed. | 108 | definition | 1.000 | https://www.icmr.gov.in/icmrobject/custom_data/pdf/downloadable-books/Anthrax_SOP.pdf | If incineration is not possible deep burial may be a viable option. | 9 | page=9,block=1 | 0.850 | valid |
| What is the purpose of the spin column protocol? | used for extracting DNA from all samples | DNA from tissues and aspirates/fluids or cultures is extracted using the DNeasy Blood & Tissue Kit or Blood Mini kit (Qiagen) as per manufacturer's instructions after the pre-treatment procedure The QIAamp DNA Stool (Qiagen) protocol is used to extract DNA from lysozyme treated stool. The spin column protocol is used for extracting DNA from all samples. DNA purity is measured using 260/280 nm in a Thermo Scientific™ NanoDrop 2000 and the ratio is expected to be above 1.5. | 314 | definition | 1.000 | https://www.icmr.gov.in/icmrobject/custom_data/pdf/downloadable-books/Anthrax_SOP.pdf | The spin column protocol is used for extracting DNA from all samples. | 23 | page=23,block=1 | 0.850 | valid |
| What does Gram-staining reveal about the appearance of certain bacteria? | bamboo stick appearance, i.e. long chain of gram-positive bacilli with non-bulging spores | Gram-staining : Reveals bamboo stick appearance, i.e. long chain of gram-positive bacilli with non-bulging spores (appears as empty space) Spores : They can be demonstrated using special stains, such as hot 5% malachite green(Ashby's method)or 0.25% sulphuric acid used in acid fast staining for spores. Spores appears as pink in colour bacilli appears as blue. Catalase Test : · Catalase produce should be done inside the petri plate Gelatin Liquefaction Test: · Slow liquefaction of gelatin Growth occurs as “Inverted fir tree appearance”(due to liquefaction of gelatin which occurs maximum at the surface, and then slows down towards the bottom) Nitrate Test: · Nitrate are reduced to nitrite. Carbohydrate fermentation Test: · Glucose, Maltose,Sucrose,Trehalose dextrin - fermented without gas. · Salicin not fermentated. | 24 | definition | 1.000 | https://www.icmr.gov.in/icmrobject/custom_data/pdf/downloadable-books/Anthrax_SOP.pdf | Gram-staining : Reveals bamboo stick appearance, i.e. long chain of gram-positive bacilli with non-bulging spores (appears as empty space) Spores : They can be demonstrated using special stains, such as hot 5% malachite green(Ashby's method)or 0.25% sulphuric acid used in acid fast staining for spores. |
| What does the appearance of a red color within a few minutes signify in the Nitrate Reduction Test? | the presence of nitrite and hence the ability of the organism to reduce nitrate to nitrite. | Reading and interpretation : A red color developing within a few minutes indicates the presence of nitrite and hence the ability of the organism to reduce nitrate to nitrite. E.g. Enterobacteriaceae If red color does not develop, a pinch of zinc powder is added and shaken. If a red color develops on addition of zinc, it confirms that the test was negative. E.g. Erwinia spp If no color develops following addition of Zinc, it confirms that nitrite was further reduced to nitrogen. E.g. Ps. aeruginosa. · · · · | 83 | definition | 1.000 | https://www.icmr.gov.in/icmrobject/custom_data/pdf/downloadable-books/Anthrax_SOP.pdf | Reading and interpretation : A red color developing within a few minutes indicates the presence of nitrite and hence the ability of the organism to reduce nitrate to nitrite. E.g. | 33 | page=33,block=0 | 0.700 | valid |
| What does the growth control well contain during the procedure? | broth with inoculum, no antimicrobial | Overnight culture of test and control organisms 5. Required antibiotic in powder form (either from the manufacturer or standard laboratory accompanied by a statement of its activity in mg/unit or per ml. Clinical preparations should not be used for reference technique) 6. Required solvent for the antibiotic 7. Sterile distilled water - 500 ml 8. Suitable nutrient broth medium (cation adjusted Muller Hinton Broth Medium). 9. Micropipettes 20 – 200 µl, 10 µl with sterile tips Procedure: 1. Prepare antimicrobial stock and serial two-fold dilution of the antimicrobial range to be tested in broth as in macrobroth dilution. 2. Add 0.1 ml of the antibiotic into the wells 3. Include one sterility well (only broth, no antimicrobial/inoculum) and one growth control well (broth with inoculum, no antimicrobial) in each tray 4. Inoculum preparation: a. Prepare 0. | 772 | definition | 1.000 | https://www.icmr.gov.in/icmrobject/custom_data/pdf/downloadable-books/Anthrax_SOP.pdf | Include one sterility well (only broth, no antimicrobial/inoculum) and one growth control well (broth with inoculum, no antimicrobial) in each tray 4. | 16 |
| What indicates gelatinase activity in the gelatin cultures? | Liquefication of gelatin i.e. absence of setting of gelatin at low temperature | Reading and interpretation : Keep the gelatin cultures in a beaker of ice or in the refrigerator before taking reading. Liquefication of gelatin i.e. absence of setting of gelatin at low temperature indicates gelatinase activity. E.g. S. aureus, Pseudomonas spp. Positive S. | 120 | definition | 1.000 | https://www.icmr.gov.in/icmrobject/custom_data/pdf/downloadable-books/Anthrax_SOP.pdf | Reading and interpretation : Keep the gelatin cultures in a beaker of ice or in the refrigerator before taking reading. Liquefication of gelatin i.e. absence of setting of gelatin at low temperature indicates gelatinase activity. | 32 | page=32,block=0 | 0.700 | valid |
| What is molecular typing used for in epidemiological studies? | to trace the source of infection | PCR with specific primers can be used for further confirmation. Molecular Typing: It is used for epidemiological studies to trace the source of infection. Various methods available are as follows : · MLVA (Multiple locus variable number of tandem repeat analysis) · AFLP ( Amplified fragment length polymorphisms) | 121 | definition | 1.000 | https://www.icmr.gov.in/icmrobject/custom_data/pdf/downloadable-books/Anthrax_SOP.pdf | Molecular Typing: It is used for epidemiological studies to trace the source of infection. | 22 | page=22,block=2 | 0.700 | valid |
| What is the definition of MIC in the context of microdilution wells? | the lowest concentration of antimicrobial agent that completely inhibits growth of the organism in the microdilution wells as detected by the unaided eye | Reading of plates: 1. Check purity of inoculum 2. Check sterility control well. It should be clear/non-turbid 3. Check growth control for adequate growth of at least 2mm button formation 4. The MIC is the lowest concentration of antimicrobial agent that completely inhibits growth of the organism in the microdilution wells as detected by the unaided eye 5. When single skipped well is seen, read the highest MIC 6. Do not report results if more than one skipped well is present 7. For trimethoprime and sulfonamides, read the end point at the concentration in which there is = 80% reduction in growth as compared to the control Recording template for microbroth dilution MIC: Fig : Broth microdilution test | 201 | definition | 1.000 | https://www.icmr.gov.in/icmrobject/custom_data/pdf/downloadable-books/Anthrax_SOP.pdf | The MIC is the lowest concentration of antimicrobial agent that completely inhibits growth of the organism in the microdilution wells as detected by the unaided eye 5. | 17 | page=17,block=0 |
| What is the formula used to calculate the Antibody Index in an ELISA test? | dividing the O.D. value of each sample by cut-off value | Calculate the Ab (Antibody) Index of each determination by dividing the O.D. value of each sample by cut-off value. Validation criteria: All 3 criteria given below have to be satisfied for every ELISA run before declaring test results. The three validation criteria to be met are: 1. The O.D. of the Calibrator should be >0.250. 2. The Ab index for Negative control should be <0.9. 3. The Ab Index for Positive control should be >1.2. Interpretation: <0. No detectable antibody to PA IgG by ELISA 0.91. Borderline positive. Follow-up testing is recommended if clinically indicated. >1. Indicative of vaccination, current or previous Anthrax infection. Quality assurance: · Kit controls (PC & NC) should give expected results · Results of the in-house QC and split sample testing should be within range. | 59 | definition | 1.000 | https://www.icmr.gov.in/icmrobject/custom_data/pdf/downloadable-books/Anthrax_SOP.pdf | Calculate the Ab (Antibody) Index of each determination by dividing the O.D. value of each sample by cut-off value. | 15 | page=15,block=1 | 0.700 |
| What is the function of Safranine Stain in bacterial differentiation? | to differentiate Gram positive and Gram negative organisms. | Acetone 100% Safranine Stain : Safranine 0.34 gm Absolute alcohol/rectified spirit 10.0 ml Distilled water 90.0 ml Add absolute alcohol to the powder kept in a mortar. Grind the paste into a smooth paste. Add distilled water and mix well Filter through filter paper into a stoppered bottle. Use: Widely used in diagnostic bacteriology to differentiate Gram positive and Gram negative organisms. Quality Control : Positive control : a heat fixed smear of S. aureus, Gram positive violet/purple cocci in clusters. Negative control : a heat fixed smear of E. coli, Gram negative pink/red bacilli. | 335 | definition | 1.000 | https://www.icmr.gov.in/icmrobject/custom_data/pdf/downloadable-books/Anthrax_SOP.pdf | Use: Widely used in diagnostic bacteriology to differentiate Gram positive and Gram negative organisms. Quality Control : Positive control : a heat fixed smear of S. | 28 | page=28,block=2 | 0.700 | valid |