CN114946720A - Method for determining acute and chronic toxicity of cephalexin to brachionus calyciflorus - Google Patents
Method for determining acute and chronic toxicity of cephalexin to brachionus calyciflorus Download PDFInfo
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Abstract
The invention relates to the field of aquatic ecology organisms and discloses a method for determining acute and chronic toxicity of cephalexin to brachionus calyciformis. Collecting calyx flower brachionus, performing monoclonal cloning, and culturing the larva in a culture container; selecting cefalexin with the purity of more than or equal to 98 percent, and preparing the cefalexin into test liquid according to a mother liquid dilution method and EPA; selecting rotifers in the culture containers, respectively placing the rotifers in the culture containers under different test liquid mass concentration gradients, repeatedly testing each concentration gradient, and observing and recording the rotifers in each culture container. The influence of cefalexin with different concentrations on the feeding behavior and the population dynamics of the rotifers can be known, and meanwhile, the ecotoxicology effect of water pollutants on the rotifer population dynamics can be known. Therefore, the manual work can timely and effectively use and manage the antibiotics in the water ecology according to the measured water ecology data, so that the water ecology system keeps corresponding balance, and scientific guidance is provided for accurate and safe use of the cephalosporin antibiotics.
Description
Technical Field
The invention relates to the field of aquatic ecology organisms, and particularly relates to a method for determining acute and chronic toxicity of cephalexin to brachionus calyciformis.
Background
Antibiotics are secondary metabolites produced by microorganisms in the life process and derivatives thereof, can effectively inhibit the growth, metabolism or death of sensitive strains at low concentration, and can be extracted from a microorganism culture solution or chemically synthesized for clinical use. Antibiotic drugs have played an important role in agriculture, animal husbandry and in the treatment of infectious diseases.
Cephalosporins are antibiotics that began to be used in the 50 s of the 20 th century; as a first generation oral cephalosporin, cephalexin is one of the most abundant classes of antibiotics consumed by humans; at present, cephalosporin antibiotics have been developed from the first generation to the fourth generation, nearly 70 cephalosporin antibiotics are developed and marketed globally, and about 46 cephalosporin antibiotics are clinically applied in China; the sales of cephalosporin antibiotics worldwide increases at a rate of 10% on average, accounting for more than 40% of the world's sales of anti-infective drugs.
With the wide use and even abuse of antibiotics in the human medical field, pathogenic bacteria carrying antibiotic resistance genes are increasing, and constitute serious threat to human health, even a plurality of 'super bacteria' appear. Long-term, large-amount and continuous discharge can cause 'false persistent' pollution of water environment antibiotics, becomes a new pollutant in the environment, and has toxic and harmful effects on aquatic organisms and human health. Studies show that the toxic effect of antibiotics on aquatic organisms is greatly different, the half-maximal effect concentration (EC50) is generally in the order of mg/L, for example, 48h acute toxicity EC50 of oxolinic acid on Daphnia magna is 4.6mg/L, the aerocin is 40mg/L, some antibiotics EC50 are higher than 200mg/L, the oxytetracycline is even as high as 1000mg/L, and the luminescent bacterium Vibrio Fischeri (Vibrio Fischer) is sensitive to antibiotics in the mu g/L range. Experiments show that norfloxacin has high acute toxicity to daphnia magna, the death rate of norfloxacin is obviously increased along with the increase of norfloxacin concentration and the prolongation of exposure time, norfloxacin with concentration lower than sub-lethal concentration obviously changes the population growth mode of daphnia magna, and the instantaneous growth rate and the population size are reduced; sulfonamides, chloramphenical, quinolones and tetracyclines can inhibit the reproduction of daphnia magna to different degrees.
Rotifers are zooplankton widely distributed in various water bodies, and have become one of important tested organisms in the fields of aquatic ecotoxicology and water environment monitoring due to the characteristics of rapid propagation, short generation time, sensitivity to changes of environmental factors and the like.
However, research shows that more than 80% of antibiotic drugs are not completely absorbed and converted and are directly discharged out of the body along with urine, feces and the like, and finally enter the ecological environment. The existing treatment technology of the sewage treatment plant can not effectively remove the active ingredients, and part of the undegraded active ingredients are finally discharged into the natural water body along with the treated sewage. Aqueous environments contain a large number of microorganisms which are tolerant to contaminants but beyond their tolerance limits exhibit toxic effects and even death.
In the existing water ecosystem, the inhibition effect of cefalexin under different concentrations on rotifers cannot be known, so that corresponding remedial measures cannot be made manually, and the balance of the water ecosystem is influenced.
Disclosure of Invention
In order to solve the technical problems, the invention provides a method for determining acute and chronic toxicity of cephalexin to brachionus calyciformis.
In order to achieve the purpose, the technical scheme of the invention is as follows:
a method for determining acute and chronic toxicity of cephalexin to brachionus calyciflorus comprises the following steps:
collecting calyx flower brachionus, performing monoclonal cloning, and culturing the larva in a culture container;
selecting cefalexin with the purity of more than or equal to 98 percent, and preparing the cefalexin into test liquid according to a mother liquid dilution method and EPA;
and selecting the rotifers in the culture containers, respectively placing the rotifers in the culture containers under different mass concentration gradients of the test liquid, repeatedly testing each concentration gradient, and observing and recording the condition of the rotifers in each culture container.
As a preferable scheme of the invention, the culture container contains culture solution adopting EPA formula and bait, wherein the bait is Scenedesmus obliquus cultured by HB-4 culture medium, and after centrifugal concentration, the culture container is fed with 1.0 x 10 per day 6 cells/mL。
In a preferred embodiment of the present invention, the cultivation environment conditions of the cultivation container are 25 ℃, 130lx of illumination intensity, and 14 h: 10h of day length ratio L: D.
As a preferable scheme of the invention, the test liquid adopts distilled water to prepare 1.0g/L mother liquor, and is placed in a refrigerator at 4 ℃ for standby.
In a preferred embodiment of the present invention, the culture vessel contains 6 different concentration gradients of the test liquid, which are 31.25mg/L, 62.5mg/L, 125mg/L, 250mg/L, 500mg/L and 1000mg/L, respectively, and a culture vessel without the test liquid is provided.
As a preferred embodiment of the present invention, 5mL of the test liquid and 1.0X 10 in density are placed in each of the culture vessels 6 The Scenedesmus obliquus, 10 rotifers with an age of < 4h were placed in the culture vessel and each concentration gradient was repeated 3 times for 24 h.
In a preferred embodiment of the present invention, the culture vessel contains 6 different concentration gradients of the test liquid, which are 0mg/L, 0.5mg/L, 1.5mg/L, 4.5mg/L, 13.5mg/L and 40.5mg/L, respectively, and another culture vessel is provided without the test liquid.
As a preferred embodiment of the present invention, 5mL of the test liquid and 1.0X 10 in density are placed in each of the culture vessels 6 The Scenedesmus obliquus is cultured in the culture container by 10 rotifers with age of less than 4h, and then placed in a constant-temperature illumination incubator with temperature of 25 ℃, illumination intensity of 130lx and day length ratio L: D of 14 h: 10h, and each concentration gradient is tested for 3 times and 20 h.
In a preferred embodiment of the present invention, the culture vessel contains 6 different concentration gradients of the test liquid, which are 0mg/L, 0.5mg/L, 1.5mg/L, 4.5mg/L, 13.5mg/L and 40.5mg/L, respectively, and a culture vessel without the test liquid is provided.
As a preferred embodiment of the present invention, 10mL of the test liquid and 1.0X 10 in density are placed in each of the culture vessels 6 The Scenedesmus obliquus is characterized in that 10 rotifers with the age of less than 4h are placed in the culture container, then the rotifers are placed in a constant-temperature illumination incubator with the temperature of 25 ℃, the illumination intensity of 130lx and the day length ratio L: D of 14 h: 10h, the test is repeated for 3 times in each concentration gradient, the algae food deposited in the culture container is suspended once every 12h, and 50% of the test liquid is replaced every 24 h.
In conclusion, the invention has the following beneficial effects: the influence of cefalexin with different concentrations on the feeding behavior and the population dynamics of the rotifers can be known, and meanwhile, the ecotoxicology effect of water pollutants on the rotifer population dynamics can be known. Therefore, the manual work can timely and effectively use and manage the antibiotics in the water ecology according to the measured water ecology data, so that the water ecology system keeps corresponding balance, and scientific guidance is provided for accurate and safe use of the cephalosporin antibiotics.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below, it is obvious that the drawings in the following description are only some embodiments of the present invention, and for those skilled in the art, other drawings can be obtained according to the drawings without creative efforts.
FIG. 1 is a schematic flow chart of the method for determining acute and chronic toxicity of cephalexin to brachionus calyciformis.
FIG. 2 is a schematic diagram showing the relationship between the death rate of Brachypodium calycinum and the concentration of cephalexin.
FIG. 3 is a schematic diagram showing the relationship between the feeding rate of Brachystemma calycinum and the concentration of cephalexin.
FIG. 4 is a schematic diagram showing population densities of Brachionus calycinus at different concentrations of cephalexin.
FIG. 5 is a schematic diagram of population growth parameters ABCDEF of Brachionus calycinus at different concentrations of cephalexin.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
The Brachypodium calycinum used in the examples of the present invention was collected from the specular lake of Wen lake of turnip lake, Anhui, and the cephalexin used in the experiments was purchased from Shanghai Aladdin Biotechnology, Inc.
The embodiment of the invention is shown in figure 1, and discloses a method for determining acute and chronic toxicity of cephalexin to brachiocephalus septempunctata, which comprises the following steps:
s100, culturing rotifers, namely collecting calyx flower brachiocephalus, performing monoclonal cloning, and culturing larvae in a culture container.
The culture container contains culture solution and bait prepared from EPA, the bait is Scenedesmus obliquus cultured in HB-4 culture medium, and is fed with 1.0 × 10 per day after centrifugal concentration 6 cells/mL, and the cultivation environmental conditions are that the temperature is 25 ℃, the illumination intensity is 130lx, and the ratio of the day length L to the day length D is 14h to 10 h. The culture time is more than 1 month. Before the experiment, the calyx flower brachiocephalus rotifer is pre-cultured for two weeks in the behavior period, so that the rotifer population is in an exponential growth state.
S200, preparing a test solution, namely selecting cefalexin with the purity of more than or equal to 98%, preparing the cefalexin by a mother solution dilution method, preparing 1.0g/L of mother solution by using distilled water before an experiment, and placing the mother solution in a refrigerator at 4 ℃ for later use. This was formulated with EPA at the time of the experiment as the test liquid at the desired concentration.
S300, testing the rotifers, namely selecting the rotifers in the culture containers, respectively placing the rotifers in the culture containers under different test liquid mass concentration gradients, repeatedly testing each concentration gradient, and observing and recording the condition of the rotifers in each culture container.
Specifically, in the acute toxicity test of the rotifers, 10 rotifers with the age of less than 4h in a culture container are selected firstly and are respectively placed in culture containers under different test liquid mass concentration gradients, specifically 31.25mg/L, 62.5mg/L, 125mg/L, 250mg/L, 500mg/L and 1000mg/L, and a culture container without the test liquid is additionally arranged as a control group.
In each of the above culture vessels, 5mL of the test liquid and a density of 1.0X 10 were placed 6 The scenedesmus obliquus is provided with 3 repeated tests in each test solution concentration group, and 21 groups are counted. Observing and recording the survival number of rotifers in each culture container after 24h of experiment, and obtaining 24hLC of the cephalexin to brachial calyx by adopting a probability unit method 50 The value is obtained.
Under the conditions of 25 ℃ and illumination intensity of 130lx, 1 rotifer is killed when the concentration of cephalexin is 31.25mg/L, and all rotifer death begins to appear when the concentration of cephalexin is 1000mg/L, wherein, the relationship between the mortality and the cephalexin concentration is shown in a figure 2, wherein, y represents the mortality, and x represents the logarithm of the cephalexin concentration with a base of 10; the 24hLC50 value of the cephalexin to calyx flower brachionus larva obtained by a probability unit method is 144.11mg/L, and the 95% confidence limit is 112.851-191.862 mg/L.
In the feeding experiment of the rotifers, 10 rotifers with the age of less than 4h in a culture container are selected firstly and are respectively placed in culture containers under different test liquid mass concentration gradients, specifically 0mg/L, 0.5mg/L, 1.5mg/L, 4.5mg/L, 13.5mg/L and 40.5mg/L, and a culture container without the test liquid is additionally arranged as a control group.
In each of the above culture vessels, 5mL of the test liquid and a density of 1.0X 10 were placed 6 The Scenedesmus obliquus is placed in a constant-temperature illumination incubator with the temperature of 25 ℃, the illumination intensity of 130lx and the day length ratio L: D of 14 h: 10h, 3 repeated tests are set for each test solution concentration group, 21 groups are counted, and after the experiment is carried out for 20h, 1mL of solution is extracted from each culture container and used for counting algae cells and calculating the feeding rate of rotifer.
As shown in fig. 3, according to the experimental analysis, the data are in accordance with the normal distribution P of 0.2 > 0.05, and the homogeneity of variance test qualification P of 0.61 > 0.05, and the single-factor analysis of variance can be used, and the single-factor analysis of variance shows that the concentration of cephalexin has a significant influence P of < 0.05 on the feeding rate of brachionus calyciflorus. The multiple comparison results show that the feeding rate of rotifers is obviously inhibited along with the increase of the concentration of the test solution, except that the concentration groups of 1.5mg/L and 4.5mg/L have no obvious difference.
In the population growth experiment of the rotifers, 10 rotifers with the age of less than 4h in a culture container are selected and respectively placed in culture containers under different test liquid mass concentration gradients, specifically 0mg/L, 0.5mg/L, 1.5mg/L, 4.5mg/L, 13.5mg/L and 40.5mg/L, and a culture container without test liquid is additionally arranged as a control group.
In each of the above culture vessels, 10mL of the test liquid and a density of 1.0X 10 were placed 6 Then placing the culture container in a constant-temperature illumination culture box with the temperature of 25 ℃, the illumination intensity of 130lx and the day length ratio L: D of 14 h: 10h, setting 3 repeated tests for each test solution concentration group, totaling 21 groups, suspending algae food deposited in the culture container once every 12h, replacing 50% of test solution every 24h, observing and recording the number of various female bodies and dormant eggs of Brachionus calycinus in each treatment group, and replacing the counted rotifers into the culture container again. The experiment continued until the population began to decline and was terminated.
The population density changes of Brachionus calycinus under different concentrations of cephalexin are shown in FIG. 4. At each concentration, the population density of brachionus calyciflorus gradually increases with the passage of time, the experimental group of 1.5-40.5mg/L reaches the maximum density at the 15 th day, and the control group and the experimental group of 0.5mg/L reach the maximum density at the 16 th day and the 17 th day respectively. The results of single-factor analysis of variance and multiple comparisons show that when the concentration of cephalexin is 0.5mg/L, the maximum density of the brachionus calyciformis population is not obviously different from that of the control group.
As shown in FIG. 5, population density of the 1.5-40.5mg/L treated group decreased faster than that of the control group in the later period of the experiment due to influence of cephalexin, so that there was a significant decrease P < 0.05 in the average population density among the cephalexin treated groups, but there was no significant difference among the three treated groups of 1.5mg/L, 4.5mg/L and 13.5mg/L, while cephalexin at 40.5mg/L minimized the rotifer average population density, as shown in FIG. 5-B.
In addition, the cephalexin with the concentration of 1.5mg/L and above obviously inhibits the population growth rate of Brachypodium calycinum, the population growth rate of rotifer is gradually reduced along with the increase of the cephalexin concentration, but no obvious difference exists between the 1.5mg/L and 4.5mg/L, and no obvious difference exists between the three cephalexin concentrations of 4.5mg/L, 13.5mg/L and 40.5mg/L, as shown in a figure 5-C.
The single-factor variance analysis of the yield of the wheel worm offspring shows that: when the concentration is more than or equal to 1.5mg/L, the yield of the offspring is obviously inhibited by P < 0.05, while the experimental groups of 1.5mg/L, 4.5mg/L and 13.5mg/L have no obvious difference, and the 40.5mg/L experimental group is obviously reduced by P < 0.01 compared with the low concentration group, as shown in figure 5-E.
Results of analysis on resting eggs of rotifers show that the 0.5mg/L cefalexin treatment group has no significant difference from the control group, when the concentration of cefalexin is more than or equal to 1.5mg/L, the yield of resting eggs is significantly improved, and the resting eggs of the 13.5mg/L treatment group are the highest in significance, as shown in FIG. 5-D.
When the concentration is more than or equal to 4.5mg/L, the average cross-mixing rate of the rotifer population is remarkably improved by P < 0.05, but when the concentration is 13.5mg/L, the population has the highest cross-mixing rate of 0.1973 +/-0.0254, as shown in figure 5-F.
According to the results of the resting egg yield and the mixed crossing rate, the cephalexin has obvious influence on the sexual reproduction of the brachionus calyciformis.
In the experiment, the algae cells are counted by using a high-power microscope, and the feeding rate of the rotifers is calculated according to the following equation:
G＝v/n×(lnC t -lnC tf )/t×(C tf -C 0 )/(lnC tf -lnC 0 )
wherein G is the feeding rate, V is the volume of the test solution, t is the feeding time, n is the number of rotifers per test group, C 0 And C tf Initial and final algal densities, C, of the addition test liquid culture vessel, respectively t Final algal density for the control group of culture vessels without added test liquid.
The population growth rate is as follows: r ═ lnN t -lnN 0 )/t
Wherein N is 0 Is the initial rotifer density, i.e. 2ind./ml, N t The rotifer population density at t days after the experiment, t being the number of days the experiment was performed. According to Dumont&According to the method of Sarma, 4-6 data are selected to calculate the population growth rate when the rotifer population is in an exponential growth period, and finally the average value of each experimental group is obtained.
The following is an explanation of the definitions in fig. 5:
maximum population density: in the experimental process, the maximum density of the brachionus calyciformis population per unit volume can be reached.
Yield of resting eggs: the total number of resting eggs laid by the rotifers per day in 10mL of culture medium.
Yield of offspring: and in unit time and unit volume, generating total post-algebra by the brachionus calyciformis population, and calculating the area between a total female body growth curve and an X axis by referring to methods of Snell & Hoff and the like.
Average mixing rate: the ratio of the mixed females to the total females in the population.
In the embodiment of the invention, the feeding behavior of zooplankton is often influenced by environmental factors, especially residual pollutants in the environment, the concentration of cephalexin has obvious influence on the feeding rate of Brachionus calyciformis, and the feeding rate of Rotiformis is obviously inhibited along with the increase of the concentration of the test solution.
In the examples of the invention, we found that feeding rate of rotifers is obviously reduced under high concentration cefalexin treatment, because cefalexin inhibits activity of rotifer protein phosphatase, thereby influencing nervous system of rotifers and swimming behavior thereof, and finally leading feeding behavior of rotifers to be inhibited. Under the cefalexin treatment, the swimming speed of the rotifer may be kept unchanged, but the feeding behavior of the rotifer is finally hindered because the movement speed of cilia on the rotifer mouth organ is inhibited by antibiotics.
In the embodiment of the invention, the concentration of cephalexin has obvious influence on the population density, the population growth rate, the yield of resting eggs and the number of offspring of brachionus calyciflorus. With the increase of the concentration of the cephalexin being more than or equal to 1.5mg/L, the maximum population density, the average population density, the population growth rate and the offspring yield of the rotifer are all obviously reduced compared with a control group and a 0.5mg/L treatment group, but the maximum population densities among the 1.5mg/L-40.5mg/L treatment groups are not obviously different; the average population density and offspring production between the three treatment groups of 1.5mg/L, 4.5mg/L and 13.5mg/L were not significantly different, while the two population parameters were reduced to the minimum at a concentration of cephalexin of 40.5 mg/L. In addition, the cephalexin with the concentration of 1.5mg/L and above obviously inhibits the population growth rate of Brachypodium calycinum, the population growth rate of rotifer is gradually reduced along with the increase of the antibiotic concentration, but no obvious difference exists between the 1.5mg/L and 4.5mg/L, and no obvious difference exists between the three cephalexin concentrations of 4.5mg/L, 13.5mg/L and 40.5 mg/L. Therefore, cephalexin has a remarkable inhibiting effect on the population density and population growth rate of brachionus calyciflorus, the reproductive rate of the brachionus calyciflorus is reduced by shortening the service life of the brachionus calyciflorus, the population growth rate and population density of the brachionus calyciflorus are changed, the number of individuals carrying non-mixed eggs by the brachionus calyciflorus is reduced, and the population fertility is inhibited.
In the embodiment of the invention, the yield analysis of the resting eggs of the rotifer shows that the 0.5mg/L cefalexin treatment group has no obvious difference with the control group, when the concentration of cefalexin is more than or equal to 1.5mg/L, the yield of the resting eggs is obviously improved, and the resting eggs of the 13.5mg/L treatment group are obviously the highest; when the concentration is more than or equal to 4.5mg/L, the average mixing rate of the rotifer population is obviously improved, and the population has the highest mixing rate when the concentration is 13.5 mg/L. As can be seen from the results of the resting egg yield and the mixed crossing rate, the cephalexin obviously improves the sexual reproduction of the branchionus calyciformis, but the survival rate of the rotifer is extremely low due to the cephalexin concentration of 40.5mg/L at the highest concentration, so the sexual reproduction rate is reduced.
With increasing concentrations of environmental toxicants, the total female rotifer production was gradually suppressed, while the mixing rate was significantly increased, thereby increasing the production of mixed females forming resting eggs per unit volume, but with further increasing concentrations, the high mortality rate decreased the production of mixed females producing resting eggs.
In the embodiment of the invention, the influence of different concentrations of cefalexin on the feeding behavior and population dynamics of the rotifers can be known, and meanwhile, the ecotoxicology effect of water pollutants on the rotifer population dynamics can be known. Therefore, the manual work can timely and effectively use and manage the antibiotics in the water ecology according to the measured water ecology data, so that the water ecology system keeps corresponding balance, and scientific guidance is provided for accurate and safe use of the cephalosporin antibiotics.
The previous description of the disclosed embodiments is provided to enable any person skilled in the art to make or use the present invention. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the generic principles defined herein may be applied to other embodiments without departing from the spirit or scope of the invention. Thus, the present invention is not intended to be limited to the embodiments shown herein but is to be accorded the widest scope consistent with the principles and novel features disclosed herein.
Claims (10)
1. A method for determining acute and chronic toxicity of cephalexin to brachionus calyciflorus is characterized by comprising the following steps:
collecting calyx flower brachionus, performing monoclonal cloning, and culturing the larva in a culture container;
selecting cefalexin with the purity of more than or equal to 98 percent, and preparing the cefalexin into test liquid according to a mother liquid dilution method and EPA;
and selecting the rotifers in the culture containers, respectively placing the rotifers in the culture containers under different mass concentration gradients of the test liquid, repeatedly testing each concentration gradient, and observing and recording the rotifer condition in each culture container.
2. The method for determining acute and chronic toxicity of cephalexin on brachionus calycinus as claimed in claim 1, wherein the culture container contains culture solution and bait with EPA formula, the bait is Scenedesmus obliquus cultured in HB-4 culture medium, and the bait is fed by 1.0 x 10 per day after centrifugal concentration 6 cells/mL。
3. The method for determining acute and chronic toxicity of cephalexin to brachionus calycinus as claimed in claim 1, wherein the cultivation environment conditions of the cultivation container are 25 ℃, the illumination intensity is 130lx and the day length ratio L: D is 14 h: 10 h.
4. The method for determining acute and chronic toxicity of cephalexin on brachionus calycinus as claimed in claim 1, wherein the test liquid is prepared into 1.0g/L mother liquid with distilled water, and is placed in a refrigerator at 4 ℃ for standby.
5. The method for determining acute and chronic toxicity of cephalexin on brachionus calycinus as claimed in claim 1, wherein the culture container contains 6 different concentration gradients of the test liquid, respectively 31.25mg/L, 62.5mg/L, 125mg/L, 250mg/L, 500mg/L and 1000mg/L, and a culture container without the test liquid is provided.
6. The method for determining acute and chronic toxicity of cephalexin on Brachypodium calycinum as claimed in claim 5, wherein 5mL of the test liquid and a density of 1.0X 10 are placed in each of the culture containers 6 The Scenedesmus obliquus, 10 rotifers with an age of < 4h were placed in the culture vessel and each concentration gradient was repeated 3 times for 24 h.
7. The method for determining acute and chronic toxicity of cephalexin on brachionus calycinus as claimed in claim 1, wherein the culture container contains 6 different concentration gradients of the test liquid, which are 0mg/L, 0.5mg/L, 1.5mg/L, 4.5mg/L, 13.5mg/L and 40.5mg/L, and a culture container without the test liquid is provided.
8. The method for determining acute and chronic toxicity of cephalexin on brachionus calycinus as claimed in claim 7, wherein 5mL of the test liquid and the density of 1.0X 10 are placed in each of the culture containers 6 The Scenedesmus obliquus is grown into 10 old Scenedesmus obliquus<4h of rotifers were placed in the culture vessel and then placed in a thermostated light incubator at 25 ℃, a light intensity of 130lx and a day length ratio L: D of 14 h: 10h, and the test was repeated 3 times for 20h per concentration gradient.
9. The method for determining acute and chronic toxicity of cephalexin on brachionus calycinus as claimed in claim 1, wherein the culture container contains 6 different concentration gradients of the test liquid, which are 0mg/L, 0.5mg/L, 1.5mg/L, 4.5mg/L, 13.5mg/L and 40.5mg/L, and a culture container without the test liquid is provided.
10. The method for determining acute and chronic toxicity of cephalexin on brachionus calycinus as claimed in claim 9, wherein 10mL of the test liquid is put in each of the culture containers and the density is 1.0X 10 6 The Scenedesmus obliquus is characterized in that 10 rotifers with the age of less than 4h are placed in the culture container, then the rotifers are placed in a constant-temperature illumination incubator with the temperature of 25 ℃, the illumination intensity of 130lx and the day length ratio L: D of 14 h: 10h, the test is repeated for 3 times in each concentration gradient, the algae food deposited in the culture container is suspended once every 12h, and 50% of the test liquid is replaced every 24 h.
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2005143329A (en) * | 2003-11-12 | 2005-06-09 | Japan Science & Technology Agency | Method for asepticizing zoo- and phytoplankton and method for culturing rotifer using the same method for asepticization |
| CN102692478A (en) * | 2012-05-31 | 2012-09-26 | 北京师范大学 | Method for detecting toxicity of PFOS (perfluorooctane sulfonate) in water through rotifers |
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2022
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2005143329A (en) * | 2003-11-12 | 2005-06-09 | Japan Science & Technology Agency | Method for asepticizing zoo- and phytoplankton and method for culturing rotifer using the same method for asepticization |
| CN102692478A (en) * | 2012-05-31 | 2012-09-26 | 北京师范大学 | Method for detecting toxicity of PFOS (perfluorooctane sulfonate) in water through rotifers |
Non-Patent Citations (1)
| Title |
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| 项尚飞: "头孢氨苄对萼花臂尾轮虫的急慢性毒性测定方法", 《2021年安徽省青少年科技创新大赛》 * |
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