Appendix XI J Microbial Limit Test Method
The tests for microbial limit are designed for non-sterile products and their APIs, excipients to determine the microbial contamination, including tests for bacteria count, mold count and the test for specified micro-organisms.
The microbial limit test should be performed in the 100-Class clean area with one-direction air flow introduced in, which settled in the 10000-Class clean environment. And all the tests should strictly comply with sterile technique to protect recontamination. The cleanliness of the one-direction air flow area, working bench and the environment should be validated based upon current national criteria on Test Method for Airborne Particles, Airborne Microbe and Settling Microbe in Clean Room (area) of the Pharmaceutical Industry regularly.
If the substances to be examined have surface-active agents, neutralizing agents or inactivators, their efficacy and absence of influence for micro-organisms must be demonstrated.
The culturing temperature for bacteria is 30~35?C; for molds and yeasts 23~28?C; and for specified micro-organisms 35~37?C,unless otherwise prescribed.
The results should be reported in 1g, 1ml, 10g, 10ml or 10cm2.
Test Amount
Test amount is the amount of the sample to be examined once (in g, ml or cm2).
The test amount is normally 10g or 10ml, unless otherwise prescribed for chemical films 100 cm2; for expensive or micro-packaged drugs, reduced test amount is acceptable. The sample to be examined for Salmonella another 10g or 10ml is required.
While testing, sample from more than 2 minimum packages, for films at least 4 sheets.
Generally speaking, randomly sample 3 times of the test amount (more than two minimum packages).
Preparation of Test Solution
The suitable method for test solution preparation depends upon the physical and biological characteristics of the substance. If the preparation introduces water bath for heating, the temperature shouldn’t exceed 45?C. As soon as the test solution is prepared, the culture medium should be added within 1 hour.
Unless otherwise prescribed, the common preparation methods are presented as follows.
1.Liquid products
Take 10ml of the sample, add in sterile buffered sodium chloride-peptone solution pH 7.0 to 100ml, and mix well to obtain 1 in 10 dilution. For oils, add in suitable quantity of sterile polysorbate 80 to evenly-disperse the test solution; and for water-soluble liquid products, mixed sample concentrate can be used as test solution.
2.Solid, semisolid or viscous products
Take 10g of the sample, add in sterile buffered sodium chloride-peptone solution pH 7.0 to 100ml, and mix well with homogenizer or other suitable methods, to obtain 1 in 10 dilution. Add in suitable quantity of sterile polysorbate 80 and properly heat with water bath to disperse the sample evenly.
3.Products prepared by special procedures
(1) Water-insoluble products
Method 1 Take 5g (or 5ml) of the sample, and add to the beaker in which there is dissolved sterile mixture(temperature not more than 45?C) containing 5g of Tween 80, 3g of glyceryl monostearate and 10g of polysorbate 80. Stir to be mass with sterile glass rod. Then slowly add in 45?C sterile buffered sodium chloride-peptone solution pH 7.0 to 100ml, while stirring. Emulsify completely to obtain 1 in 20 dilution.
Method 2 Take 10g of the sample, and add to the suitable container in which there are 20ml of sterile isopropyl myristate (refer to the Sterility Test in Appendix XI H Sterility Test for preparation methods) and sterile glass balls. If necessary, increase the amount of isopropyl myristate used. Shake and vibrate to dissolve completely. Then add in 45?C sterile buffered sodium chloride-peptone solution pH 7.0 to 100ml, vibrate for 5~10 minutes to extract, and stand still to laminate the water and oil. Take the water layer as 1 in 10 dilution.
(2) Films products
Take 100 cm2 of the sample, cut into pieces, add in 100ml of sterile buffered sodium chloride-peptone solution pH 7.0 (If necessary increase the amount added), soak, and vibrate to obtain 1 in 10 dilution.
(3) Enteric or colonic targeting products
Take 10g of the sample, add in sterile phosphate buffer solution pH 6.8 ( for enteric targeting products) or sterile phosphate buffer solution pH 7.6 ( for colonic targeting products) to 100ml, and place in 45?C water bath. Shake and vibrate to dissolve, and obtain 1 in 10 dilution.
(4) Aerosol and Spray products
Take specified amount of the sample, freeze in freezing room for about 1hour. Take out and sterilize the open part of the sample quickly. Drill a small hole with sterile steel awl in the sterilize part, and settle in room temperature. Gently rotate the container to release all the propellants slowly. Then suck all th products out with sterile syringe, add to the suitable quantity of sterile buffered sodium chloride-peptone solution pH 7.0 (If water-insoluble component exists, add in suitable quantity of sterile polysorbate 80), and mix well. Take the sample equals to contain 10g or 10ml of the products, dilute to obtain 1 in 10 dilution
(5) Products with antimicrobial activity
If the products to be examined have antimicrobial activity, the test should be performed after removing the activity. Common methods following:
①Culture medium dilution
Transfer specified amount of the test solution to a great quantity of culture medium to reduce the sample content per unit volume, till the antimicrobial activity is removed. When performing bacteria, molds and yeasts count, take 2ml of test solution with the same dilution level. Transfer each 1ml of the test solution to different dishes with equal quantity, pour in agar culture medium, mix well, allow to solidify, incubate and count. The sum of the number of Colony forming Units from each dish with the 1 ml test solution transferred to is namely the CFU per ml test solution. Calculate the average CFU for each ml, and report CFU following the counting rule of plate-count method; When performing the test for specified micro-organisms, increase the amount of enrichment medium used.
②Micro-organisms accumulating by Centrifuging and precipitating
Take a quantity of the test solution, centrifuge at 3000r /min for 20 minutes (if precipitate exists, centrifuge at 500r/min for 5 minutes firstly, and then take the supernatant to further centrifuge), discard the supernatant, take 2ml of the accumulated solution at the bottom, and add in diluent to original volume.
③Membrane filtration
Refer to the Membrane filtration in the Section of Bacteria, Molds and Yeasts Counts.
④Neutralization
For the products containing mercurial, arsenics or antiseptics which inhibit micro-organisms, proper neutralizing agents or inactivating agents can be chose to remove their antimicrobial activity. The neutralizing agents or inactivating agents
can add in the diluent or culture medium used.
Bacteria, Molds and Yeasts Count
Validation of the Count Methods
While establishing the test methods of the microbial limit for the product, the validation of the bacteria, molds and yeasts count methods should be performed to assure that the applied method is suitable for testing the bacteria, molds and yeasts in the product. If the components in product or the test conditions are changed, which will possibly influence the test results, the method should be revalidated.
Perform the validation following the methods and requirements in Preparation of Test Solution and Bacteria, Molds and Yeasts Count. For each of the micro-organism listed, separate tests for recovery should be performed.
Strains
The strain used to validate are not more than 5 passages (the freeze-dry strain removed from the microbiological culture collection center is defined as 0 generation), and maintained using suitable culture maintenance techniques to ensure their biological characteristics.
Escherichia coli [CMCC (B) 44 102]
Staphylococcus aureus [CMCC (B) 26 003]
Bacillus subtilis[CMCC (B) 63 501]
Candida albicans[CMCC (F) 98 001]
Aspergillus niger[CMCC (F) 98 003]
Preparation of Test Strains
Inoculate the Nutrient broth or Nutrient Agar with the fresh culture materials of Escherichia coli, Saphylococcus aureus, Bacillus subtilis , and incubate for 18~24 hours; Inoculate the Modified Martin Broth or Modified Martin Agar with the fresh culture materials of Candida albicans, and incubate for 24~48 hours. Use 0.9% sterile sodium chloride solution, prepare the test suspension containing 50~100 CFU per ml with the above culture materials. Inoculate the Modified Martin Agar slant with the fresh culture materials of Aspergillus niger, and incubate for 5~7 days. Add in 3~5ml of 0.9% sterile sodium chloride solution to wash out the spores. Then suck the spores suspension (use the sterile capillary with thin sterile cotton or gauze at its port to filter mycelium) to sterile test tube, and prepare the test suspension containing 50~100 CFU per ml using 0.9% sterile sodium chloride solution.
Validation Method
At least three separate parallel tests should be performed for validation, and calculate
the recovery for each micro-organism for each test respectively.
(1)Test group
Plate count method: take 1ml of test solution with the possible minimum dilution level and 50~100 CFU of test micro-organism, add to the dish respectively, immediately pour agar medium in. 2 parallel dishes are prepared for each micro-organism, and calculate the CFU following the plate-count method. Membrane filtration method: take specified quantity of test solution with the possible minimum dilution level, filter, rinse and add 50~100 CFU of micro-organism into the final diluent used to rinse, filer, and calculate the CFU following the membrane filtration method.
(2)Micro-organism group
To determine the number of CFU for the test micro-organism added.
(3)Test control group
Take specified quantity of test solution, and determine the number of CFU in test solution.
(4)Diluent control group
If dispersing,emulsifying, neutralizing, centrifuging or membrane filtering are introduced in the preparation of test solution, diluent control group should be added to observe the influence on the micro-organisms. Use the chosen diluent in place of test solution, add in test micro-organism to obtain the final strain concentration of 50~100 CFU per ml. Determine the number of CFU with the same preparation methods and CFU count methods as the test group.
Interpretation of the Results
In the three separate parallel tests, the recovery for diluent control group (the percentage for mean counts of diluent control group of the results obtained with micro-organism group) should be all not less than 70%. If the recovery for test group (mean counts of test group minus the results obtained with test control group, and the percentage for this value of the results from micro-organism group) should be all not less than 70%, the test solution preparation methods and count methods can be used to test bacteria, molds and yeasts counts in product; If any result obtained from test group is less than 70%, Culture medium dilution, Micro-organisms accumulating by Centrifuging and precipitating, Membrane filtration, Neutralization or a combination of the above measures should be used to remove the antimicrobial activity, and revalidate the methods.
The validation can be performed while the bacteria, molds and yeasts counts are testing.
Test Methods
The test methods include plate-count method and membrane filtration method.. When testing, use the validated count method to test the bacteria, molds and yeasts counts.
Take the well-mixed test solution prepared with validated methods, and dilute with sterile buffered sodium chloride-peptone solution pH 7.0 to obtain 1 in 10, 1 in 102 and 1 in 103 dilutions.
1. Plate-count method
Using this method, 2~3 suitable test solutions with consecutive dilution level should be chosen.
For dishes 90mm in diameter, add to the sterile dish 1ml of the test solution and 15~20ml of Nutrient Agar Medium or Rose Bengal Medium or Yeast Extract Peptone Dextrose Agar Medium at not more than 45?C, mix well, allow to solidify and incubate upside down. Prepare for each medium at least 2 dishes for each level of dilution.
Negative Control
Add to the sterile dish 1ml of the chosen diluent and the medium, allow to solidify and incubate upside down. Prepare for each medium 2dishes. There must be no growth of micro-organisms.
Incubating and Counting
Unless otherwise prescribed, incubate the bacteria for 48 hours, count the CFU every day, and normally report the results at 48 hours; incubate the molds and yeasts for 72 hours, count the CFU every day, and normally report the results at 48 hour; prolong the incubating period to 5~7 days if necessary, count and report then. The plates on which the colonies grow up and appear laminar are unsuitable to count. After counting, calculate the mean number of CFU for each dilution level, and report the results following the rule. If the numbers of CFU on the two plates for the same dilution level are not less than 15, the difference between plates shouldn’t be more than 1 time.
Generally, Nutrient Agar Medium is used for bacteria counting; Rose Bengal Medium for molds and yeasts; Yeast Extract Peptone Dextrose Agar Medium for yeasts. Under special conditions, if colonies of molds and yeasts are detected on Nutrient Agar Medium, or colonies of bacteria on Rose Bengal Medium, they are counted separately. And then compare the colonies number of molds and yeasts detected on Nutrient Agar Medium, or colonies of bacteria on Rose Bengal Medium to that of molds and yeasts on Rose Bengal Medium or bacteria on Nutrient Agar Medium, and take the higher results.
For the liquid products containing honey, bee milk, use Rose Bengal Medium to determine molds, and Yeast Extract Peptone Dextrose Agar Medium to determine yeasts, and the calculate the sum.
CFU Count Reporting Rule
It is acceptable to chose the dilution level with 30~300 mean CFU of bacteria, yeasts, 30~100 of molds to be the criteria to report the results (keep two effective numbers).
(1) If the number of CFU count for only one dilution level conforms to the previous described criteria, multiply the mean number of CFU count by dilution factor to report the results.
(2) If the numbers of CFU counts for two dilution levels conform to the previous described criteria, the results depend on the ratio (multiply the number of CFU count by dilution factor and then calculate the quotient of the value from higher dilution level to that from lower one). If the ratio is not more than 2, multiply the numbers of CFU counts by dilution factor, and then calculate the average as the reported results. If the ratio is more than 2 but not more than 5, multiply the number of CFU count obtained from the lower dilution level by dilution factor to report the results. If the ratio is more than 5, or the colonies from higher dilution level are more or equal to that from lower one, which is abnormal, firstly find out the cause and then perform the testing. Revalidate the methods if necessary.
(3) If the mean number of CFU count for each dilution level is less than 30, multiply the number of CFU count obtained from the lowest dilution level by dilution factor to report the results.
(4) if no growth of micro-organisms on the plate for each dilution level, or only the growth on the plate for lowest dilution level but the mean number of CFU count is less than 1, multiply <1 by dilution factor to report the results.
2. membrane filtration method
Use membrane filters having a nominal pore size not greater than 0.45μm, and diameter about 50mm. The type of filter material is chosen such that the bacteria-retaining efficiency is not affected by the components of the sample to be examined and the diluent chosen. Before using, sterilize the filters and membranes properly. The integrality of the used membrane before and after filtering should be ensured. For the water-soluble sample, filter a small quantity of diluent to moisten the membrane. For oil sample, the membranes and filters should be dry enough. To obtain the maximum filtering efficiency of the membrane, please ensure that the test solution and diluent cover the whole surface of the membrane. After filtering test solution, rinse the membrane with diluent. Each time 100ml of diluent is used for each membrane. The total filtration amount for each membrane should be controlled to protect the micro-organisms from injury.
Take the quantity of test solution which equals to 1g or 1ml of sample on each filtration membrane, add to suitable quantity of diluent, mix well and filter. If the micro-organisms in 1g or 1ml of sample is too much, choose the 1ml of the test solution with suitable dilution level, and filter. Rinse the filtration membrane with sterile buffered sodium chloride-peptone solution pH 7.0, the methods should follow the related section in Validation. After rinsing, transfer the membrane to the Nutrient Agar Medium or Rose Bengal Medium or Yeast Extract Peptone Dextrose Agar Medium plates with the micro-organisms side up. Then incubate. For each medium, at least one membrane is used.
Negative Control
Take 1ml of the chosen diluent, perform the testing as previous described for membrane filtration as negative control. There must be no growth of micro-organisms.
Incubating and Counting
Incubating conditions and counting methods are the same as the plate-count methods. Not more than 100 CFU on each membrane.
CFU Count Reporting Rule
Report the results for the number of CFU in 1g or 1ml of the sample; if there is no growth on the membrane, report as <1(for each membrane, filer 1g or 1ml of the sample), or <1 multiplied by the dilution factor.
Test for Specified Micro-organisms
Validation of Test Method for Specified Micro-organisms
While establishing the test methods of the microbial limit for the product, the validation of test for specified micro-organism should be performed to assure that the applied method is suitable for testing the specified micro-organism in the product. If the components in product or the test conditions are changed, which will possibly influence the test results, the method should be revalidated.
While validating, select the corresponding validating strain according to the specified micro-organism listed in the criteria of Microbial Limit. For example, to validate coliform group Escherichia coli is used as validating strain. Validation should be performed according to the rules of test solution preparation and specified micro-organism test methods and the following requirements.
Strains
The requirements is the same as that in the validation of Bacteria, Molds and Yeasts Count method.
Escherichia coli [CMCC(B) 44 102]
Staphylococcus aureus [CMCC (B) 26 003]
Salmonella paratyphi B [CMCC (B) 50 094]
Pseudomonas aeruginosa [CMCC (B) 10 104]
Clostridium sporogenes [CMCC (B) 64 941]
Preparation of Test Strains
Inoculate the Nutrient broth or Nutrient Agar Medium with the fresh culture materials of Escherichia coli, Staphylococcus aureus, Salmonella paratyphi B, Pseudomonas aeruginosa, and Fluid Thioglycollate Medium with the fresh culture materials of Clostridium sporogenes. Incubate for 18~24 hours; Use 0.9% sterile sodium chloride solution, prepare the test suspension containing 10~100 CFU per ml.
Validation Method
(1)Test group
Take specified quantity of the test solution and 10~100 CFU test micro-organism, add to the enrichment medium, determine according to the corresponding test methods for specified micro-organism. Using membrane filtration method, take specified quantity of test solution, filter, rinse and add micro-organism into the final diluent used to rinse, filer, then add in enrichment medium or take out the membrane and transfer to enrichment medium.
(2)Negative micro-organism control group
The purpose is to validate the specificity of the method. Detailed procedures are the same as test group, to validate E. coli , Coliform, Salmonella, S. aureus is used as negative control; to validate S. aureus, P. aeruginosa, Clostridium, E. coli is used. There must be no growth of the negative micro-organisms.
Interpretation of the Results
There must be no growth of the negative micro-organisms in the negative micro-organism control group. If there is growth in the test group, the test solution preparation and specified micro-organism test methods can be performed; if there is no growth, Culture medium dilution, Micro-organisms accumulating by Centrifuging and precipitating, Membrane filtration, Neutralization or a combination of the above measures should be used to remove the antimicrobial activity, and revalidate the methods.
The validation can be performed while the specified micro-organisms are testing. Test methods
Test for specified micro-organism should be performed with the validated methods, and the actual used quantity of enrichment medium should be also according to the validation.
Positive Control Test
Positive control should be tested, while performing the test for specified micro-organisms in the products. The inoculums’volume is 10~100 CFU, the procedures are the same as the test for specified micro-organisms. There must be growth in positive control test.
Negative Control Test
Take 10ml of the diluent, perform the testing as test for specified micro-organisms. There must be no growth in negative control test.
(1) Escherichia coli
Take 10ml of the test solution (equal to 1g, 1ml, 10cm2of the sample), inoculate directly or after treating to Lactose Bile Medium, and incubate for 18~24hours. Prolong to 48 hours if necessary.
Inoculate 0.2ml of above culture materials to the test tube containing 5ml of MUG medium, incubate, and observe under 366nm ultraviolet light at 5-hour, 24-hour. Meanwhile use the MUG medium without inoculation as control. If the medium show fluorescence, record as MUG positive; or MUG negative. After observation, add several drops of Indole TS along the tube wall, if the rosy red appears, record as Indole positive; the original color of the TS remains, record as Indole negative. The results for control must be MUG negative and Indole negative.
MUG positive and Indole positive, indicate the presence of E. coli in the sample; MUG negative and Indole negative indicate the absence of E. coli; if MUG positive and Indole negative, or MUG negative and Indole positive, inoculate and scratch the culture materials from Lactose Bile Medium to plate with Eosin Methylene Blue Agar or MacConkey agar medium, and incubate or 18~24 hours.
If there is no growth of micro-organism on the plate, or the colonies characteristic on the plate not conform to those listed in Table 1, then indicate the absence of E. coli in the sample. If the characteristic conform to or are similar to those listed in Table 1, isolating, purification, dyeing for microscope observation and other suitable biochemical tests should be used to confirm if the micro-organism is E. coli.
(2) Coliform
Take three tubes containing suitable quantity (not less than 10ml) of Lactose Bile Medium, add in 1ml of 1 in 10 diluted test solution, 1 in 100 diluted test solution (containing 0.01g or 0.01ml of sample), 1 in 1000 diluted test solution (containing 0.001g or 0.001ml of sample) respectively, add 1ml of diluent into another Lactose Bile Medium tube as negative control. Incubate for 18~24 hours.
If no growths of micro-organism in Lactose Bile Medium tube, or the growths of micro-organism without acid and gas released can indicate the absence of Coliform; if acid and gas can be detected, inoculate and scratch the culture materials from tubes to the plates with Eosin Methylene Blue Agar or MacConkey agar medium, and incubate or 18~24 hours.
If there is no growth of micro-organism on the plate, or the colonies characteristic on the plate not conform to those listed in Table 2 or it is belong to Non- Gram-negative no-spores rod bacteria, then indicate the absence of E. coli in the sample. If the characteristic conform to or are similar to those listed in Table 1, and be determined as Gram-negative rod, the confirmation test should be performed.
Confirmation Test
Provoke 4~5 doubted colonies from the isolation medium as previous described, inoculate to Lactose Fermentation Tubes, and incubate for 24~48hours. If acid and gas can be detected, can indicate the presence of Coliform, or the absence of Coliform.
Based upon the tubes number detected Coliform, and report the Coliform CFU numbers in 1g or 1ml of the sample following Table 3.
(3) Salmonella
Transfer 10g or 10ml of sample, directly or after treating to the suitable quantity (not less than 200ml) of Nutrient broth , mix well with homogenizer or other suitable method, and incubate for 18~24 hours.
Inoculate 1 ml of the above culture materials to 10ml of TTB medium, and incubate for 18~24houre. Then separately inoculate and scratch to the plated with DHL (or SS ) Medium and MacC (or EMB) Medium, and incubate for 18~24 hours ( prolong to 40~48 hours if necessary). If there is no growth of micro-organism on the plate, or the colonies characteristic on the plate not conform to those listed in Table 4, then indicate the absence of Salmonella.
If the characteristic conform to or are similar to those listed in Table 4, with inoculating needle provoke 2~3 colonies and to TSI deep slant, both inoculate to the surface and puncture in the deep slant. Incubate for 18~24 hours, if no red appears on the slant and no yellow appears at the bottom; or yellow appears on the slant and no black appears at the bottom, then indicate the absence of Salmonella in the sample. Otherwise, perform suitable biochemical tests and serum coagulation tests to confirm if the micro-organism is Salmonella.
(4) Pseudomonas aeruginosa
Take 10ml of the test solution (equal to 1g, 1ml, 10cm2of the sample), inoculate directly or after treating to suitable quantity (not less than 100ml) of Lactose Bile Medium, and incubate for 18~24hours. Inoculate and scratch the above culture
materials to the plate with Cetrimide Agar Medium and incubate for 18~24hours.
The typical colony for Pseudomonas aeruginosa is flat, amorphous, the edge is diffused and the surface is moist, grey-white. There is blue-green pigment diffused around. If there is no growth of micro-organism on the plate, or the colonies characteristic on the plate not conform to those listed above, then can indicate the absence of Pseudomonas aeruginosa in the sample. If the characteristic conform to or are similar to those listed above, provoke 2~3 colonies and inoculate to Nutrient Agar slant, and incubate for 18~24 hours. Take the slant culture materials, Gram-dye, observe by microscope and perform Oxidase test.
Oxidase Test
Place clean filter paper into the plate. Take the slant culture materials and spread on the paper. Drip fresh-prepared 1 % N,N-Dimethyl-p-Phenylenediamine Dihydrochloride TS. If the culture materials appear pink and then turn purple red gradually, recognize as Oxidase test positive, or the negative.
If the materials is the non Gram negative no-spores rod bacteria or the Oxidase Test shows negative, then can indicate the absence of Pseudomonas aeruginosa in the sample. Otherwise Pyocyanin Test should be performed.
Pyocyanin Test
Inoculate the slant culture materials on the slant with the PDP agar medium, incubate for 24 hours, add 3~5ml of Chloroform into the tube, stir medium to pieces, shake and mix well. Stand still for a while, transfer the chloroform phase into another tube, add in 1ml of 1 mol/L hydrochloric acid, shake, stand still for a while and observe. If the hydrochloric acid solution appears pink, recognize as the Pyocyanin test positive, or the negative. Meanwhile prepare a slant of PDP agar medium without inoculation with the same methods as negative control, of which the results should be negative.
If the above doubted micro-organism is Gram-negative rod, and both the Oxidase test and Pyocyanin test shows positive, then can indicate the presence of Pseudomonas aeruginosa. If the above doubted micro-organism is Gram-negative rod, and both the Oxidase test and Pyocyanin test shows negative, then suitable biochemical tests should be performed to confirm if the micro-organism is Pseudomonas aeruginosa. (5) Staphylococcus aureus
Take 10ml of the test solution (equal to 1g, 1ml, 10cm2of the sample), inoculate directly or after treating to suitable quantity (not less than 100ml) of Sodium (Potassium) Tellurite broth (or Nutrient broth) Medium, and incubate for 18~24hours, prolong to 48hours if necessary. Inoculate and scratch the above culture materials to the plate with Egg Yolk Salt Agar Medium (or Mannitol Salt Agar Medium) and incubate for 24~72hours. If there is no growth of micro-organism on the plate, or the colonies characteristic on the plate not conform to those listed in Table 5, then indicate the absence of Staphylococcus aureus in the sample.
If the characteristic conform to or are similar to those listed in Table 5, provoke 2~3 colonies and inoculate to Nutrient Agar slant, and incubate for 18~24 hours. Take the slant culture materials, Gram-dye, inoculate in Nutrient Broth Medium and perform Plasma Coagulase test.
Plasma Coagulase test
Take 3 sterilized testing tubes, add in 0.5ml of the mixture of plasma and sterile water (1:1), then add in 0.5ml of the Nutrient Broth with doubted micro-organism (or the strain suspension prepared with the Nutrient Agar slant culture materials), 0.5ml of he Nutrient Broth with Staphylococcus aureus (or the strain suspension prepared with the Nutrient Agar slant culture materials), 0.5ml of Nutrient broth or 0.9% sterile Sodium Chloride solution, which namely the test tube, the positive control tube and the negative control tube. Incubate these three tubes in the meantime; observe after 3 hours to 24 hours. The plasma in negative control tube should flow smoothly, the one in positive control tube should coagulate, and if the plasma in test tube coagulate recognize as Plasma Coagulase Test positive, or the negative. If the results of positive or negative control tubes do not conform to the criteria, prepare plasma and perform the test again.
If the above doubted micro-organism is non Gram positive coccus, and the plasma coagulase test shows negative, can indicate the absence of Staphylococcus aureus in the sample.
(6) Clostridium
Take 10ml of the test solution (equal to 1g, 1ml of the sample), two portions. Maintain one of the portions at 80?C for 10 minutes and cool down quickly. Then inoculate the two portions of the test solution directly or after treating to 100ml of 0.1% fresh Cooked Meat Medium. Incubate the medium tubes under anaerobic conditions for 72~96hours. If there is no turbid, gas given off, meat disappeared, smelly gas and other appearances, can indicate the absence of the Clostridium. Otherwise, inoculate 0.2ml of the above culture materials on the plate with Columbia Agar Medium containing Gentamycin, and incubate under anaerobic conditions for 48~72hours. If there is no growth of micro-organism on the plate, can indicate the absence of Clostridium in the sample; if there is growth of micro-organism on the plate, provoke 2~3 colonies, Gram dye and perform Catalase Test.
Catalase Test
Place the colonies on the above plates onto the clean glass slide, and drip 3% Hydrogen peroxide TS, if there are bubbles appearing on the surface of the colony recognize as Catalase Test positive, or the negative.
If the above doubted micro-organism is Gram-positive Clostridium, with or without oval or global spores, and the Catalase test shows negative, can indicate the presence of Clostridium; or, indicate the absence of the Clostridium in the sample.
Interpretation of the Results
When the presence of specified micro-organism or other pathogens is detected in the sample, the results is valid on this time, no need to test again.
If any item of the bacteria, molds and yeasts counts in the sample do not conform to the specification, sample at random from the same batch, perform the test again separately for two times. Report the CFU number with the mean value of the three results.
If the molds and yeasts are detected in the ocular products for eye, there must be no growth of micro-organism for the results from the two times of re-test, only which can indicate that the molds and yeasts counts numbers conform to the criteria for the products.
If the test results for bacteria, molds and yeasts and specified micro-organisms all conform to the criteria for the products, can indicate the products conform to the specification; any item out of criteria will indicate that the products out of specification.
Diluents
After the diluent is prepared, sterilize with the validated procedures.
1. Sterile Buffered Sodium Chloride-peptone Solution pH 7.0
Refer to Sterility Test (Appendix XI H).
2. Sterile Phosphate Buffer Solution pH 6.8 or Sterile Phosphate Buffer Solution pH 7.6
Prepare following Buffer (Appendix XV D), filter, discharge and sterilize.
3. Sterile 0.9% Sodium Chloride Solution
Weigh 9.0g of Sodium Chloride, dissolve and add water to 1000ml, filer, discharge and sterilize.
Culture Medium and Preparation Methods
Prepare the culture medium according to the following formulas, or use the dehydrated medium produced following the formulas. After preparing, sterilize with the validated procedures.
1. Nutrient Agar Medium, Nutrient Broth Medium, Fluid Thioglycollate Medium, Modified Martin Broth or Modified Martin Agar Medium
Refer to Sterility Test (Appendix XI H).
2. Rose Bengal Agar
Peptone 5.0 g
Glucose 10.0 g
Potassium dihydrogen phosphate 1.0 g
Magnesium sulfate 0.5 g
Rose Bengal sodium 0.0133 g
Agar 14.0 g
Water 1000 mL
Mix all the ingredients except for glucose and rose bengal sodium, dissolve with gentle heat and filter, then add glucose and rose bengal sodium, repackage and sterilize.
3. Yeast Peptone Dextrose Agar (YPD)
Peptone 10.0 g
Yease 5.0g
Glucose 20.0 g
Agar 14.0 g
Water 1000 mL
Mix all the ingredients except for glucose, dissolve with gentle heat and filter, then add glucose, repackage and sterilize.
4. Bile Lactose Broth (YPD)
Peptone 20.0 g
Lactose 5.0 g
Sodium chloride 5.0 g
Dipotassium hydrogen phosphate 4.0 g
Potassium dihydrogen phosphate 1.3 g
Ox bile salt 2.0g
(or sodium deoxycholate) (0.5 g)
Water 1000 mL
Mix all the ingredients except for lactose and ox bile salt or sodium deoxycholate, dissolve with gentle heat, adjust the pH to 7.4 ± 0.2 after sterilization, boil out and filter, then add lactose and bile salt or sodium deoxycholate, repackage and sterilize.
5. Bile Lactose Broth (YPD)
Take 1000 mL of non-sterilized Bile Lactose Broth, add 25 mL of 0.04% bromcresol purple indicator, discharge into the tubes with small inverted tubes basing on the required volume and sterilize. The inverted tubes should be suitable to observer the gas releasing.
6. Eosin-Methyleneblue Agar Medium (EMB)
Nutrient agar medium 100 mL
20% lactose solution 5 mL
Eosin sodium indicating solution 2 mL
Methylene blue indicating solution 1.3 to 1.6 mL
Thaw the nutrient agar medium, cool to 60℃, add other three sterilized solutions, mix well and pour into plate.
7. MacConkey Agar Medium (MacC)
Peptone 20.0 g
Lactose 10.0 g
Ox bile salt 5.0g
Sodium chloride 5.0 g
1% neutral red indicating solution 3 mL
Agar 14.0 g
Water 1000 mL
Mix all the ingredients except for lactose, 1% neutral red indicating solution, ox bile salt and agar, dissolve with gentle heat, adjust the pH to 7.2 ± 0.2 after sterilization, add agar, thaw with heat, add other ingredients, shake well, repackage and sterilize. Cool to 60℃ and pour into plate.
8. 4-methylumbelliferyl-β-D-glu-curonide medium
Peptone 10.0 g
Manganess sulfate 0.5 mg
Zinc sulfate 0.5 mg
Magnesium sulfate 0.1 g
Sodium chloride 5.0 g
Calcium chloride 50 mg
Potassium dihydrogen phosphate (anhydrous) 0.9 g
Dipotassium hydrogen phosphate (anhydrous) 6.2 g
Sodium sulfite 40 mg
Sodium deoxycholate 1.0 g
MUG 75 mg
Water 1000 mL
Mix all the ingredients except for MUG, dissolve with gentle heat, adjust the pH to 7.3 ± 0.1 after sterilization, add MUG, repackage as 5 mL per tube and sterilize.
9. Triple Sugar Iron Agar medium
Peptone 10.0 g
Beef extract powder 5.0 g
Lactose 10.0 g
Saccharose 10.0 g
Glucose 1.0 g
Sodium chloride 5.0 g
Ferrous sulfate 0.2 g
Sodium thiosulfate 0.2 g
0.2% phenolsulfonphthalein indicating solution 12.5 mL
Agar 12.0 g
Water 1000 mL
Mix all the ingredients except for the three sugar, 0.2% phenolsulfonphthalein indicating solution and agar, dissolve with gentle heat, adjust the pH to 7.3 ± 0.1 after sterilization, add agar, thaw with heat, add other ingredients, shake well, repackage and sterilize to prepare high and low (2~3cm) short slant .
10. Tetrathionate Broth Base
Peptone 5.0 g
Ox bile salt 1.0g
Calcium carbonate 10.0 g
Sodium thiosulfate 30.0 g
Water 1000 mL
Mix all the ingredients, dissolve with gentle heat and sterilize.
Take above medium, add 0.2 mL of iodine TS and 0.1 mL of light green TS per 10mL medium, and mix well.
11. Salmonella-Shigella agar
Peptone 5.0 g
Beef extract powder 5.0 g
Lactose 10.0 g
Ox bile salt 8.5 g
Natrium citricum 8.5 g
Ferric ammoniun citrate 1.0 g
Sodium thiosulfate 8.5 g
Neutral red indicating solution 3 mL
Light green TS 0.33 mL
Agar 16.0 g
Water 1000 mL
Mix all the ingredients except for lactose, neutral red indicating solution and agar, dissolve with gentle heat, adjust the pH to 7.2 ± 0.1 after sterilization, filter, add agar
and thaw with heat, add other ingredients, shake well, repackage and sterilize. Cool to 60℃ and pour into plate.
12. Deoxycholate Hydrogen Sulfide Lactose Agar (DHL)
Peptone 10.0 g
Beef extract powder 3.0 g
Lactose 10.0 g
Saccharose 10.0 g
Sodium deoxycholate 1.0 g
Sodium thiosulfate 2.3 g
Natrium citricum 1.0 g
Ferric ammoniun citrate 1.0 g
Neutral red indicating solution 3 mL
Agar 16.0 g
Water 1000 mL
Mix all the ingredients except for sugars, indicating solution and agar, dissolve with gentle heat, adjust the pH to 7.2 ± 0.1 after sterilization, add agar and thaw with heat, add other ingredients, shake well, cool to 60℃ and pour into plate.
13. Cetrimide Agar Medium
Peptone 10.0 g
Beef extract powder 3.0 g
Sodium chloride 5.0 g
Cetrimide 0.3 g
Agar 14.0 g
Water 1000 mL
Mix all the ingredients except for agar, dissolve with gentle heat, adjust the pH to 7.5 ± 0.1 after sterilization, add agar and thaw with heat, repackage and sterilize, cool to 60℃ and pour into plate.
14. Tellurite Broth Medium
Add 0.2 mL newly prepared 1% sodium (potassium) telluite TS into 100 mL of sterilized nutrient broth medium, mix well.
15. Egg-yolk Salt Agar
Peptone 6.0 g
Beef extract powder 1.0 g
Sodium chloride 30.0 g
10% egg-yolk solution 100 mL
Agar 14.0 g
Water 650 mL
Mix all the ingredients except for 10% egg-yolk solution, dissolve with gentle heat, adjust the pH to 7.6 ±0.1 after sterilization, sterilize and cool to 60℃, add 10% egg-yolk solution with aseptic technique, shake well and pour into plate.
Preparation of 10% egg-yolk solution Take out the yolk from a fresh egg with aseptic technique, add it into 100 mL of 10% sterilized sodium chloride solution, shake well.
16. Mannitol Salt Agar
Peptone 10.0 g
Beef extract powder 1.0 g
Mannitol 10.0 g
Sodium chloride 75.0 g
Phenolsulfonphthalein indicating solution 2.5 mL
Agar 14.0 g
Water 1000 mL
Mix all the ingredients except for mannitol, phenolsulfonphthalein indicating solution and agar, dissolve with gentle heat, adjust the pH to 7.4 ± 0.2 after sterilization, add agar and thaw with heat, filter, repackage and sterilize, cool to 60℃and pour into plate.
17. Lactose Broth
Peptone 20.0 g
0.04% bromocresol purple indicating solution 25 mL
Lactose 10.0 g
Water 1000 mL
Mix all the ingredients except for 0.04% bromocresol purple indicating solution, dissolve with gentle heat, adjust the pH to 7.2 ±0.2 after sterilization, add the indicating solution, repackage into tubes with small inverted tubs as 3 mL per tube, and sterilize.
18. Pyocyanin Detection Medium (PDP)
Peptone 20.0 g
Magnesium chloride (anhydrous) 1.4 g
Potassium chloride (anhydrous) 10.0 g
Glycerin 10 mL
Agar 14.0 g
Water 1000 mL
Mix peptone, magnesium chloride, potassium chloride and water, dissolve with gentle heat, adjust the pH to 7.3 ± 0.1 after sterilization, add glycerin and agar, thaw and mix well, repackage into tubes, sterilize and prepare slants.
19. Cooked Meat Medium
Preparation of meat particles Take fresh beef, remove fat and tendon, boil with distilled water for 10 minutes. Cut into little particles of 5 mm3, weigh and add distilled water with a ratio of 1:3 (meat: distilled water), put at 4 to 10℃ for 18 to 20 hours, boil for 1 hour, filter with white cloth (filtrate is 1:3 beef dip). Wash the filter residue with tap water twice, add quantum satis of sodium hydrate solution, stir and
A/O法活性污泥中氨氧化菌群落的动态与分布 摘要: 我们研究了在厌氧—好氧序批式反应器（SBR)中氨氧化菌群落(AOB)和亚硝酸盐氧化菌群落(NOB)的结构活性和分布。在研究过程中,分子生物技术和微型技术被用于识别和鉴定这些微生物。污泥微粒中的氨氧化菌群落结构大体上与初始的接种污泥中的结构不同。与颗粒形成一起，由于过程条件中生物选择的压力,AOＢ的多样性下降了。DGGE测序表明，亚硝化菌依然存在,这是因为它们能迅速的适应固定以对抗洗涤行为。DGGE更进一步的分析揭露了较大的微粒对更多的AOB种类在反应器中的生存有好处。在SＢR反应器中有很多大小不一的微粒共存，颗粒的直径影响这AＯB和NOB的分布。中小微粒（直径<0．6ｍm）不能限制氧在所有污泥空间的传输。大颗粒(直径>０．9mm）可以使含氧量降低从而限制NＯB的生长。所有这些研究提供了未来对ＡOB微粒系统机制可能性研究的支持。 关键词:氨氧化菌(AＯB），污泥微粒,菌落发展，微粒大小,硝化菌分布，发育多样性 ?简介 在浓度足够高的条件下，氨在水环境中对水生生物有毒，并且对富营养化有贡献。因此,废水中氨的生物降解和去除是废水处理工程的基本功能。硝化反应,将氨通过硝化转化为硝酸盐,是去除氨的一个重要途径。这是分两步组成的，由氨氧化和亚硝酸盐氧化细菌完成。好氧氨氧化一般是第一步,硝化反应的限制步骤:然而,这是废水中氨去除的本质。对16S ｒRNA的对比分析显示，大多数活性污泥里的氨氧化菌系统的跟?-变形菌有关联。然而，一系列的研究表明,在氨氧化菌的不同代和不同系有生理和生态区别,而且环境因素例如处理常量,溶解氧，盐度,pH，自由氨例子浓度会影响氨氧化菌的种类。因此,废水处理中氨氧化菌的生理活动和平衡对废水处理系统的设计和运行是至关重要的。由于这个原因,对氨氧化菌生态和微生物学更深一层的了解对加强处理效果是必须的。当今,有几个进阶技术在废水生物处理系统中被用作鉴别、刻画微生物种类的有价值的工具。目前,分子生物技术的应用能提供氨氧化菌群落的详细分类说明。
C A T T I三级笔译综合能力真题及答案解析
CATTI三级笔译综合能力考试试题及答案解析(一) 一、Vocabulary Selection（本大题15小题．每题1.0分，共15.0分。In this part, there are 20 incomplete sentences. Below each sentence, there are four words or phrases respectively marked by letters A, B, C and D. Choose the word or phrase which best completes each sentence. There is only one right answer. ） 第1题 Since writing home to their parents for money, they had lived ________hope. A in B for C on D through 【正确答案】：A 【本题分数】：1.0分 【答案解析】 固定搭配。live in hope生活在希望中；live for为……而生活，盼望；live on继续生活，以……为主食，靠……生活；live through度过，经受过；根据句意应填A。 第2题 ________get older, the games they play become increasingly complex. A Children B Children, when they C As children D For children to
翻译专业本科人才培养方案 一、培养目标 立足湖南，面向全国，将翻译专业与英语专业、翻译专业硕士有机结合，坚持翻译理论与技巧教学与实践应用并重，培养面向全球一体化、适应中国国情与区域人才市场需要的“基础厚，口径宽，能力强，素质高”的应用型翻译专门人才。 二、专业特色及实现途径 专业特色： 英语翻译专业本着“厚基础、宽口径、强能力、高素质”的培养原则，将通识教育与专业教育相结合，在“通识”中突出“专才”，打造真正适合社会的高级英语人才。本专业学生除受到基础语言技能训练外，系统地学习口译与笔译的技巧，掌握林业、工程机械、经贸等工程领域翻译的基本技巧，掌握基本的翻译理论知识；同时，学生进一步学习汉语文学知识，提高汉语应用能力；广泛了解中国文化与英语国家的文化，从而获得作为职业译员的基本素养，工作能力和研究能力。 实现途径： （1）科学合理、与时俱进的课程体系； （2）英汉双语优良师资； （3）依托我院多年来积累的高水平实践、实习基地，多年来翻译口译大赛的承办及参赛经验以及系统完善的口笔译实训、实习经验， 保证学生翻译实践技能的培养。 （4）依托我校林业优势，借我校工程机械专业优秀师资之力，为培养“专才”铺路。 三、培养要求及保障措施 培养要求：翻译专业本科学生毕业时应达到以下知识、能力和素质要求。 （1）知识要求 通识知识：翻译专业本着“厚基础、宽口径、强能力、高素质”的培养原则，将通识教育与专业教育相结合，在“通识”中突出“专才”，打造真正适合社会的高级英语人才。学生除专业技能外，将进一步学习汉语文学知识，提高汉语应用能力；广泛了解中国文化与英语国家的文化，从而获得作为职业
清酒乳杆菌(Lactobacillus sakei),弯曲乳杆菌(Lactobacillus curvatus),明串珠菌属的肠膜明串珠菌(Leuconostoc mesenteroides)和非培养的明串珠菌(Uncultured Leuconostoc sp.) 清酒乳杆菌清酒亚种(Lactobacillus sakei subsp.sakei) 弯曲乳杆菌蜜二糖亚种(Lactobacillus curvatus subsp.melibiosus) 粪肠球菌（E.faecalis）屎肠球菌（E.faecium) 鸟肠球菌(E.avium) 酪黄肠球菌(E.casseliflavus) 坚忍肠球菌(E.durans) 鸡肠球菌E.galinarum) 芒地肠球菌(E.mundii) 恶臭肠球菌(E.maladoratum) 希拉肠球菌(E.hirae) 孤立肠球菌（E.solitarius） 棉子糖肠球菌(E.raffinosus) 假鸟肠球菌(E.pseudoavium) 粪肠球变异株(E.faecalis var)。 Abiotrophia adjacens 毗邻贫养菌 Abiotrophia defectiva 软弱贫养菌 Achromobacter spp 无色杆菌属某些种 Acinetobacter /Pseudomonas spp 不动杆菌/假单胞菌属某些种 Acinetobacter baumannii 鲍氏不动杆菌 Acinetobacter calcoaceticus 醋酸钙不动杆菌 Acinetobacter haemolyticus 溶血不动杆菌 Acinetobacter johnsonii 约氏不动杆菌 Acinetobacter junii 琼氏不动杆菌 Acinetobacter lwoffii 鲁氏不动杆菌 Acinetobacter radioresistens 抗辐射不动杆菌 Acinetobacter spp 不动杆菌属某些种 Acinetobacter spp/Pseudomonas spp 不动杆菌属某些种/假单胞菌属某些种 Acinetobacter/Pseudomonas spp 不动杆菌/假单胞菌属某些种 Actinobacillus actinomycetemcomitans 伴放线放线杆菌 Actinomyces israelii 衣氏放线菌 Actinomyces meyeri 麦氏放线菌 Actinomyces naeslundii 内氏放线菌 Actinomyces neuii anitratus 纽氏放线菌无硝亚种 Actinomyces neuii neuii 纽氏放线菌纽氏亚种 Actinomyces neuii radingae 纽氏放线菌罗亚种 Actinomyces neuii turicensis 纽氏放线菌图列茨亚种 Actinomyces odontolyticus 龋齿放线菌 Actinomyces viscosus 粘放线菌
CATTI三级笔译综合能力考试试题及答案解析(三) 一、Vocabulary Selection（本大题20小题．每题1.0分，共20.0分。In this part, there are 20 incomplete sentences. Below each sentence, there are four words or phrases respectively marked by letters A, B, C and D. Choose the word or phrase which best completes each sentence. There is only one right answer. ） 第1题 Marketing is ______ just distributing goods from the manufacturer to the final customer. A rather than B other than C bigger than D more than 第2题 The magician picked several persons ______ from the audience and asked them to help him with the performance. A by accident B at random C on occasion 第3题 English language publications in China are growing in volume and ______. A circulation B rotation C circumstance D appreciation 第4题
Microscope 显微镜 Antony van leeuwenhoek 吕文虎克 Louis Pasteur 巴斯德 Joseph Lister 李斯德 Robert Koch 郭霍 Koch postulate 郭霍法则 molecular Koch postulate 分子郭霍法则Bacterium 细菌 Coccus 球菌 Diplococcus 双球菌 Bacillus 杆菌 Spiral bacterium 螺型菌 Vibrio 弧菌 Spirillum 螺菌Polymorphism 多形性 Cell wall 细胞壁Peptidoglycan 肽聚糖Mucopeptide 粘肽 Glycopeptide 糖肽 Murein 胞壁质 N-acetylmuramic acid ，NAM N-乙酰胞壁酸N-acetylglucosamine, NAG N-乙酰葡糖胺Diaminopimelic acid, DAP 二氨基庚二酸Teichoic acid 磷壁酸 Ribitol 核糖醇 Lipoteichoic acid, LTA 脂磷壁酸 Outer membrane 外膜Lipopolysaccharide LPS 脂多糖 Lipid A 脂质A Lysozyme 溶菌酶 Protoplast 原生质体Spheroplast 圆球体Bactrrial L form 细菌L形 Cytoplasmic membrane 细胞膜 Mesosome 中介体 Cytoplasm 细胞质 Ribosome 核糖体 Plasmid 质粒 Inclusion 内含物 Metachromatic granule 异染颗粒 Volutin 纡回体 Nuclear material 核质 Nucleoid 拟核 Capsule 荚膜 Microcapsule 微荚膜 Slime layer 黏液层 Smooth colony 光滑型菌落 Mucoid colony 黏液型菌落 Rough colony 粗糙型菌落 Flagellum 鞭毛 Pilus(fimbria) 菌毛 Common pilus 普通菌毛 Sex pilus 性菌毛 Fertility 致育性 Spore 芽孢 Endospore 内芽孢 Vegetative form 繁殖体 Spore wall 芽孢壁 Cortex 皮质层 Coat 芽孢壳 Exosporium 芽孢外衣 Light microscope 光学显微镜 Electron microscope 电子显微镜 Dark microscope 暗视野显微镜 Gram stain 革兰染色法 Bacterial metabolism 细菌代谢 Obligate aerobe 专性需氧菌 Microaerophilic bacterium 微需氧菌 Facultative anaerobe 兼性厌氧菌 Obligate anaerobe 专性厌氧菌 Superoxide dismutase SOD 超氧化物歧化酶 Catalase 触酶 Peroxidase 郭氧化物酶 Binary fission 二分裂法 Growth curve 生长曲线 Lag phase 迟晚期 Logarithmic phase 对数生长期 Stationary phase 稳定期 Decline phase 衰亡期 Autotroph 自养菌 Heterotroph 异养菌 Saprophyte 腐生菌 Permease 通透酶 Fermentation 发酵 Pyrogen 热原质 Antibiotic 抗生素 Exotoxin 外毒素 Endotoxin 内毒素 Bacteriocin 细菌素 Culture medium 培养基 Basal medium 基础培养基 Nutrient medium 营养培养基 Selective medium 选择性培养基 Different medium 鉴别培养基 Anaerobic medium 厌养培养基 Cooked meat medium 庖肉培养基 1
英语翻译三级笔译综合能力模拟试题及答案解析（13） (1/20)SECTION 1 Vocabulary Selection In the section, there are 20 incomplete sentences. Below each sentence, there are 4 choices respectively marked by letters A,B,C and D. Choose the word or phrase which best completes each sentences. There is only ONE right answer. 第1题 It discusses the major economic, institutional social and geographical ______ that need to be addressed in the appropriate introduction and use of this technology. A.fronts https://www.doczj.com/doc/7d15086011.html,ments C.facts D.aspects 下一题 (2/20)SECTION 1 Vocabulary Selection In the section, there are 20 incomplete sentences. Below each sentence, there are 4 choices respectively marked by letters A,B,C and D. Choose the word or phrase which best completes each sentences. There is only ONE right answer. 第2题 The three largest Japanese banks are ______ into the world's largest banking group. A.assimilated B.incorporated C.embodied https://www.doczj.com/doc/7d15086011.html,bined 上一题下一题 (3/20)SECTION 1 Vocabulary Selection In the section, there are 20 incomplete sentences. Below each sentence, there are 4 choices respectively marked by letters A,B,C and D. Choose the word or phrase which best completes each sentences. There is only ONE right answer. 第3题 In addition, government has acted as the provider of pump ______ funds for new applications, but this role is increasingly being called into question. A.priming B.breaking C.emerging D.omitting 上一题下一题 (4/20)SECTION 1 Vocabulary Selection In the section, there are 20 incomplete sentences. Below each sentence, there are 4 choices respectively marked by letters A,B,C and D. Choose the word or phrase which best completes each sentences. There is only ONE right answer. 第4题 The role played by supranational entities, such as the WTO, ITU and telecoms MOU bodies ______ in and regulating this environment will be examined. A.following
浅析翻译人才培养的社会需求导向 【关键词】社会需求；双语转换；培养目标；教学模式 【摘要】从分析我国翻译事业得现状动身,提出了翻译人才培养应以社会需求为导向得观点,认为翻译工作得特别性决定了翻译人才培养模式得特别性,而传统外语专业教学模式不适合于翻译人才得培养. 近年来,我国得翻译学科建设取得了长足得进展,翻译院系和翻译研究机构在一些高校相继建立.2006年春,教育部批准在部分高校(复旦大学、广州外语外贸大学和河北师范大学)试点设立本科翻译专业.2007年1月,国务院学位委员会批准设立“翻译硕士专业学位”.翻译学科体系得健全与进展,不仅是翻译事业进展得需要,也是我国改革开放政策不断深化,经济、科学与文化事业蓬勃进展得需要. 1翻译人才培养应以社会需求为导向 人才培养要习惯社会与经济进展得需求.任何一个新专业得设置,都必须以社会需求为导向,凸显“应用”特色.也确实是讲,社会需要什么人才,学校就培养什么人才.《翻译硕士专业学位设置方案》把培养目标定位在“德、智、体全面进展、能习惯全球经济一体化及提高国家国际竞争力得需要、习惯国家经济、文化、社会建设需要得高层次、应用型、专业性口笔译人才”,正是秉承了“以社会需求为导向”得理念,将人才培养与社会需求紧密结合起来.wWwcOm 翻译历来是国际交流与交往得重要桥梁和纽带.改革开放以来,我国在政治、经济、文化和科技等各个领域得对外交流与合作日益频繁,各种国际会议日益增多,国外先进得科学技术不断涌人,迫切需要将其转化为我们自己得语言去了解、汲取和掌握.另一方面,随着我国国际地位得提高,世界各地不断掀起“ 翻译工作得特别性要求翻译专业课程设置必须理论与实践相结合,学生不仅要学习翻译理论与技巧,还要在理论指导下进行翻译实践.然而,目前我国翻译方向硕士生得培养却存在着“重理论、轻实践”得咨询题,其培养目标为高校教学人员和科研人员,培养模式偏重于学术性,对翻译得专业性和应用性则不够重视.在人学考试、培养目标、课程设置、教学安排和学位论文写作等方面,基本上按照学术型人才培养模式进行得.忽视翻译操作技能得培养,就会导致翻译实践能力偏低,不利于高层次、应用型、专业性翻译人才得培养.某大学一位教师告诉笔者,他们学校有资料翻译任务,想请新近聘用得一位翻译方向硕士毕业生承担一部分,没想到这位毕业生难道拒绝参与,讲自己是搞翻译理论研究得,不擅长翻译实践.翻译理论研究对翻译学科建设无疑起了重要得推动作用,翻译理论关于翻译实践得指导作用也是不容忽视得,但仅仅明白得理论而可不能实践,如此得毕业生就无法满足社会对翻译实践人才得需求. 由于我校去年新上了翻译方向硕士点,我曾利用参加国内翻译学术会议得机会,向一些知名外语院校得教师询咨询翻译方向硕士研究生是否应开设翻译实践课,得到得回答大多是否定得.我咨询他们,学生得操作能力差该如何办?他们告诉我,用翻译项目来弥补.翻译项目实践诚然是必要得,但翻译项目并非人人都能得到,并非总是能够得到,没有项目时该如何办?笔者认为,必须为翻译方向硕士生开设一定数量得翻译实践课,以确保他们有足够多得实践机会.就我所知,北京外国语大学对学生翻译实践能力得培养比较重视.2001年,我在北外英语学院
全国翻译资格考试三级笔译综合能力讲义 第一节考试内容介绍、定语从句讲解、练习及译法 第一部分考试介绍 一、考试题型 词汇和语法部分50题25分25分钟 阅读理解50题55分75分钟 完型填空20题20分20分钟 二、考试要求 掌握本大纲要的英语词汇；掌握并能够正确运用双语语法；具备对常用问题英语文章的阅读理解能力。 三、笔译综合能力试题的基本类型 第一部分词汇和语法部分可分为三大部分：词汇选择（Vocabulary and Grammar）1-20难度大约在四级左右；词语替代（Vocabulary Selection）21-40主要找出和划横线部分相同意思的词汇，难度在4级到6级之间；改错（Error Correction）主要有词汇和语法的两种错误。 第一部分考察的内容主要为近义词的辨析、短语介词和动词词组的搭配；语法点主要包括定语从句、状语从句、名词性从句、虚拟语气、非谓语动词等等。在其后会分专题来讲解。 第二部分阅读理解共5篇，字数在每篇150-450字之间，绝大多数在250字左右，每篇有5到10题，不定量。题材广泛，选题多样。类似于四级以上难度和题型，但是和专业四级题型更加相似。 第三部分为开放性完型填空，20空，共20分。题材广泛，选题多样。难度和专业四级等同。 四、基本复习策略 综合能力课程作为三级笔译必修的课程，主要是考察学生对于英语基本知识的了解，特别是对双语习惯的掌握，为了能够更好地为实务课程打下坚实的基础，所以希望每一位同学都不要对这门课程产生掉以轻心的念头。根据个人经验，我个人推荐以下几个复习的策略：第一、要对大纲的词汇做到十分熟悉，这种熟悉不是简单地认识，而是要学会使用，特别是在没有字典的帮助之下可以迅速而准确地判断搭配。 第二、对于语法结构的重视，但凡是学翻译就必须要对语法结构有着深入的了解，这种了解不是简单地会划分句子成分，而是将每个句子如何组合，这将决定你实务中的句型翻译的关键。 第三，阅读能力要强。任何考试都是得阅读者得天下，咱们的综合能力考试也不除外。包括实务考试，如果文章内容没有弄明白，那么对文章的翻译肯定是不行的。 以上三点经验之谈，仅供参考。 定语从句的讲解、练习及译法 1．that和which用法比较： 1) which用于非限定性定语从句中。 e.g.: He said he was busy, which was not true. 他说他很忙，那是假的。 We don’t want to enter the house, which is very cold. 我们不想进房间，因为太冷了。 2) which用于介词后做宾语。 e.g.: The room of which windows are opposite to the room is large. 窗户正对着海的房间很大。 The chair in which you are sitting is made of iron.
常见微生物 界（Domain）、门(Phylum)、纲（Class）、目(Order)、科(Family)、属(Genus)、种(Species)中英文对照
界（Domain）Bacteria细菌 Archaea古生菌
门（Phylum）Proteobacteria 变形菌门 Bacteroidetes 拟杆菌门 Actinobacteria 放线菌门Gemmatimonadetes 芽单胞菌门Acidobacteria 酸杆菌门 Planctomycetes 浮霉菌门Verrucomicrobia 疣微菌门 Chloroflexi 绿弯菌门 Nitrospirae 硝化螺旋菌门 Firmicutes 厚壁菌门 Chlorobi 绿菌门 Cyanobacteria 蓝藻细菌门Fibrobacteres 纤维杆菌门 Elusimicrobia 迷踪菌门Armatimonadetes 装甲菌门 Euryarchaeota 广古菌门 Chlamydiae 衣原体 Crenarchaeota 泉古菌门 Tenericutes 无壁菌门 Spirochaetes 螺旋体属
Alphaproteobacteria 甲型（α）变形杆菌纲Gammaproteobacteria 丙型变形菌纲Betaproteobacteria β-变形菌纲Actinobacteria 放线菌门、纲Cytophagia 纤维粘网菌Gemmatimonadetes 芽单胞菌门、纲Deltaproteobacteria δ-变形菌纲Acidobacteria-6 酸杆菌门Acidimicrobiia 酸微菌纲Verrucomicrobiae 疣微菌纲 Opitutae 丰佑菌纲 Nitrospira 消化螺菌属Thermomicrobia 热微菌门 Bacteroidia 拟杆菌纲 Bacilli 杆菌 Chloroflexi 绿弯菌门Anaerolineae 厌氧绳菌纲 Clostridia 梭状芽胞杆菌Elusimicrobia 迷踪菌门Ktedonobacteria 纤线杆菌纲Thermoplasmata 热原体纲Chlamydiia 衣原体Thaumarchaeota 奇古菌门 Mollicutes 柔膜菌纲Methanomicrobia 甲烷微菌纲Holophagae 全噬菌纲Spirochaetes 螺旋体属
active immunity（主动免疫）: Immunity acquired through direct stimulation of the immune system by antigen. active transport（主动运输）:Transport of molecules against a concentration gradient (from regions of low concentration to regions of high concentration) with the aid of proteins in the cell membrane and energy from ATP. Alcohol fermentation（乙醇发酵）：is the formation of alcohol from sugar. Yeast, when under anaerobic conditions, convert glucose to pyruvic acid via the glycolysis pathways, then go one step farther, converting pyruvic acid into ethanol, a C-2 compound. aerobe（好氧微生物）: A microorganism that lives and grows in the presence of free gaseous oxygen (O2). aflatoxin（黄曲霉毒素）: From Aspergillus flavus t, a mycotoxin that typically poisons moldy animal feed and can cause liver cancer in humans and other animals. AIDS（爱滋病）: Acquired Immune deficiency syndrome. The complex of signs and symptoms characteristic of the late phase of human immunodeficiency virus (HIV) infection. Ames test（艾姆氏实验）: A method for detecting mutagenic and potentially carcinogenic agents based upon the genetic alteration of nutritionally defective bacteria anabolism（合成代谢）: The energy consuming process of incorporating nutrients into protoplasm through biosynthesis. anaerobe（厌氧微生物）: A microorganism that grows best, or exclusively, in the absence of oxygen. antibiotic（抗生素）:A chemical substance from one microorganism that can inhibit or kill another microbe even in minute amounts. antibody（抗体）: A large protein molecule evoked in response to an antigen that interacts specifically with that antigen. antigen（抗原）: Any cell, particle, or chemical that induces a specific immune response by B cells or T cells and can stimulate resistance to an infection or a toxin. antigenic determinant（抗原决定基）:The precise molecular group of an antigen that defines its specificity and triggers the immune response. antimetabolite（抗代谢物）:A substance such as a drug that competes with, substitutes for, or interferes with a normal metabolite. antiseptic（防腐剂）：A growth-inhibiting agent used on tissues to prevent infection. antiserum（抗血清）：Antibody-rich serum derived from the blood of animals (deliberately immunized against infectious or toxic antigen) or from people who have recovered from specific nfections. antitoxin（抗毒素）：Globulin fraction of serum that neutralizesa specific toxin. Also refers to the specific antitoxin antibody itself. arthrospore（节孢子）：A fungal spore formed by the septation fragmentation of hyphae. ascospore（子囊）：A spore formed within a saclike cell (ascus) of Ascomycota following nuclear fusion and meiosis. asepsis（无菌）：A condition free of viable pathogenic microorganisms. autoantibody（自身抗体）：An "anti-self antibody having an ffinity for tissue antigens of the subject in which it is formed. autoantigen（自身抗原）：Molecules that are inherently part of self but are perceived by the
2005年5月英语三级《笔译综合能力》试题 Section 1: Vocabulary and Grammar (25 points) This section consists of 3 parts. Read the directions for each part before answering the questions. The time for this section is 25 minutes. Part 1 Vocabulary Selection In this part, there are 20 incomplete sentences. Below each sentence, there are 4 choices respectively marked by letters A, B, C and D. Choose the word or phrase which best completes each sentence. There is only ONE right answer. Blacken the corresponding letter as required on your Machine-scoring ANSWER SHEET. 1. We have had to raise the prices of our products because of the increase in the cost of ______ materials. A. primitive B. rough C. original D. raw 2. With an eighty-hour week and little enjoyment, life must have been very______ for the students. A. hostile B. anxious C. tedious D. obscure 3. Whenever the government increases public services,______ because more workers are needed to carry out these services. A. employment to rise B. employment rises C. which rising employment D. the rise of employment 4. Our flight to Guangzhou was ______ by a bad fog and we had to stay much longer in the hotel than we had expected. A. delayed B. adjourned C. cancelled D. preserved 5. Container-grown plants can be planted at any time of the year, but ______ in winter. A. should be B. would be C. preferred D. preferably 6. Both longitude and latitude ______ in degrees, minutes and seconds. A. measuring B. measured C. are measured D. being measured 7. Most comets have two kinds of tails, one made up of dust, ______ made up of electrically charged particles called plasma. A. one another B. the other C. other ones D. each other 8. Good pencil erasers are soft enough not ______ paper but hard enough so that they crumble gradually when used. A. by damaging B. so that they damage C. to damage D. damaging 9. The magician picked several persons ______ from the audience and asked them to help him with the performance. A. by accident B. at random C. on occasion D. on average 10. On turning the comer, they saw the path ______ steeply. A. departing B. descending C. decreasing D. degenerating
微生物学思考题 第一章绪论 1.微生物有哪五大共性，其中最基本的是哪一个，何故？ 微生物五大共性分别是：1:体积小，面积大；2:吸收多，转化快；3:生长旺，繁殖快；4:适应强，易变异；5:分布广，种类多。其中最基本的特性是体积小，面积大。微生物是一个突出的小体积大面积系统，从而赋予它们具有不同于一切大生物的五大共性，因为一个小体积大面积系统，必然有一个巨大的营养物质吸收面、代谢废物的排泄面和环境信息的交换面，故而产生了其余四个共性。巨大的营养物质吸收面和代谢废物的排泄面使微生物具有了吸收多，转化快，生长旺，繁殖快的特点。环境信息的交换面使微生物具有适应强，易变异的特点。而正是因为微生物具有适应强，易变异的特点，才能使其分布广，种类多。 2.为什么说巴斯德和柯赫是微生物学的奠基人？（请不要简单罗列二个人的工作，而应该对他们的工作及意义进行评论） 路易·巴斯德，主要贡献：①否认了“自生说”；②初步应用免疫学，利用预防接种法治疗疾病，给人类带来幸福；③证实了发酵作用与微生物活动有关；④发明了巴氏灭菌法。④分离鉴定了引起家蚕蚕病杆菌并提出预防措施，被誊为微生物的奠基人。 罗伯特·柯赫,专门研究细菌，特别是病原菌，对微生物学有卓越贡献：①建立微生物学研究基本技术，被誉为细菌学技术之父。②证实病害的病原菌学说（柯赫法则）。 具体证实了炭疽病菌是炭疽病的病原菌。 发现了肺结核病的病原菌。 他们将微生物大的研究从形态描述推进到生理学研究阶段，揭露了微生物是造成腐败发酵和人畜疾病的原因，并建立了分离、培养、接种和灭菌等一系列独特的微生物技术，从而奠定了微生物学的基础，开辟了医学和工业微生物等分支学科。巴斯德和柯赫是微生物学的奠基人 3.微生物包括哪几大类群？ 真核：酵母菌、霉菌、原生动物、单细胞藻类 原核：真细菌(放线菌、支原体、立克次氏体、衣原体、蓝细菌)、古细菌 非细胞结构：病毒、亚病毒(类病毒、拟病毒、朊病毒) 4.名词解释：微生物、种、菌株、型。 A 微生物：是指一切体形微小，单细胞或个体结构简单的多细胞，甚至没有细胞结构 的低等生物的统称 B种：是一个基本分类单位；是一大群表型特征高度相似、亲缘关系极其接近，与同属内其他种有明显差别的菌株的总称。 C菌株：表示任何由一个独立分离的单细胞繁殖而成的纯种群体及其一切后代（起源于共同祖先并保持祖先特性的一组纯种后代菌群）。因此，一种微生物的不同来 源的纯培养物均可称为该菌种的一个菌株。菌株强调的是遗传型纯的谱系。 D型：常指亚种以下的细分。当同种或同亚种内不同菌株之间的性状差异不足以分为新的亚种时，可以细分为不同的型。例如：按抗原特征的差异分为不同的血清型5.用一个具体事例说明人类与微生物的关系，为什么说微生物是人类的敌人，更是我们的朋友？ 对于人类来说，微生物有有害的，也有有益的，它们可以使人致病，也可以为人治病，可以给人类造成损失，也可以为人类创造财富。 致病的如葡萄球菌、天花病毒等，治病的如青霉素、噬菌体等，肠道中的双歧杆菌还有维护肠道正常细菌菌群平衡、抑制病原菌的生长、防止便秘、在肠道内合成维生素、氨基酸
模拟试题（三） 笔译综合能力 Section 1 Vocabulary and Grammar (60points) This section consists of 3 parts. Read the directions for each part before answering the questions. The time for this section is 25 minutes. Part 1 Vocabulary Selection In this part, there are 20 incomplete sentences. Below each sentence, there are 4 choices marked by letters A, B, C and D respectively. Choose the word or phrase which best completes each sentence. There is only ONE right answer. Blacken the corresponding letter as required on your Machine-scoring ANSWER SHEET. 1.Jupiter_________ perhaps the most important planet of the solar system. A．was B. were C. is D. will be 2. The common garden pea, also called the English pea, _________ for its edible seeds. A．grows B. is grown C. growing D. grown 3. The olfactory regions of the nose are yellow, richly moist, and_________. A．fully fatty substances B. full, fatty substances C. full of fatty substances D. fatty substances are full 4. _________, the moon is important because it is the nearest to the earth of all heavenly bodies.