CN103033630B - Kit for qualitative detection of lipovitellin of cyprinid fish and application thereof - Google Patents
Kit for qualitative detection of lipovitellin of cyprinid fish and application thereof Download PDFInfo
- Publication number
- CN103033630B CN103033630B CN201210557080.1A CN201210557080A CN103033630B CN 103033630 B CN103033630 B CN 103033630B CN 201210557080 A CN201210557080 A CN 201210557080A CN 103033630 B CN103033630 B CN 103033630B
- Authority
- CN
- China
- Prior art keywords
- lipovitellin
- goldfish
- fish
- kit
- mouse
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Fee Related
Links
Abstract
The invention discloses a kit for the qualitative detection of lipovitellin of cyprinid fish and the application thereof. The kit comprises goldfish lipovitellin products and mouse-anti goldfish lipovitellin polyclonal antibody. The method comprises the steps of firstly performing purifying to obtain goldfish lipovitellin, preparing mouse-anti goldfish lipovitellin polyclonal antiserum through immunized mice, and performing further purifying to obtain the mouse-anti goldfish lipovitellin polyclonal antibody. The kit can be used for identification and qualitative analysis of ovary of cyprinid fish and in vivo lipovitellin of larva fish, avoids traditional complex methods, such as sugar, phosphorus and fat dyeing and amino acid composition analysis, reduces financial resources, time and workload to a certain degree, and greatly improves the detection sensitivity on the basis that the work efficiency is improved, and the minimum detectability is about 100 ng/ml.
Description
Technical field
The invention belongs to aquaculture field, be specifically related to a kind of kit and application thereof of qualitative detection cyprinid fish lipovitellin.
Background technology
The growth of fish is divided into alevin stage, postlarva phase, young stage, immature phase, adult fish phase and declining period, the growth of the division in each period and fish sexual gland has direct relation, for raun, in its ovary, main material is livetin, and livetin plays an important role in the whole life cycle of fish.First, livetin is the important substance of embryonated egg, for fish embryo grow and prelarva early growth nutriment and energy source are provided.Secondly, in yolk generation, ovum small cell absorbs vitellogenin from blood, after processing, is stored in egg mother cell with the form of yolk granules and oil droplet, and the content of livetin directly affects embryo's survival and growing of prelarva.
Vitellogenin is produced by the liver of raun, arrive ovary with blood circulation, under the effect of cathepsin D, be cracked into rapidly three kinds of main livetins: lipovitellin, phosvitin and β ' component, be stored in yolk granules, wherein abundant with the content of lipovitellin.Similar to vitellogenin, lipovitellin is also a kind of typical GLP, and contains a small amount of phosphorus, in embryo's g and D process, has various biological function.In the maturation of egg mother cell, the further cracking of lipovitellin, discharges lipid, and produces free amino acid, for embryo and postlarva provide the main substrate of the synthetic raw material of protein and aerobic energy production, and provide required lipid at specific developmental stage.At present after deliberation the multiple fish lipovitellins such as haddock, verasper moseri, hybrid sturgeon in egg mother cell or embryo development procedure, change, and confirm that lipovitellin is by being degraded into free amino acid, for the growth of body early embryo and postlarva provides energy and nutriment; Lipovitellin also possesses the function that regulates osmotic pressure, and in the growth course of egg cell, buoyant egg is because absorbing moisture, volume enlarges markedly, and while reaching maturity, in buoyant egg, moisture reaches 85 ~ 95%, egg cell volume increases 3 ~ 5 times, for embryo's growth provides water for subsequent use.In this course, the increase of free aminoacid content, plays an important role for generation moisture flows into needed osmotic pressure, and meanwhile, the increase of liquid water content provides fish-egg to swim in buoyancy required in water body.(2009) and Ohkubo etc. (2006) confirmations such as Reith etc. (2001), Reading, the free amino acid in buoyant egg is mainly derived from the heavy chain of lipovitellin; In addition, lipovitellin also possesses immunologic function, in most of the cases, the fertilization of fish and growth will be carried out in water environment, and in water, often contain the pathogen that can cause various diseases, at early development, embryo and postlarva cannot be synthesized enough immunity related moleculars, also do not form lymphoid organ.Therefore,, before immune system is reached maturity, can only rely on maternal material to keep out the pathogen of invasion.Shi Xiaodong (2003) finds the growth that lipovitellin that rose need not Barb can resist multiple pathogen; Zhang J and Zhang SC (2011) research are found; rose need not Barb lipovitellin and Escherichia coli and staphylococcus aureus there is very strong binding ability; and energy can strengthen the phagocytic activity of macrophage; supposition lipovitellin can either be identified the pathogen of invasion; can bring out again macrophage digestion pathogen, thereby protection embryo and postlarva avoid disease.
Visible, the lipovitellin of researching fish has very big help to understanding fish early development situation.At present, purifying and the authentication method of multiple fish lipovitellin are reported both at home and abroad, but mostly adopt its characteristic that contains sugar, phosphorus, fat group, adopt specific stain or analyze its sugar, phosphorus, fat content to determine whether it is lipovitellin.This method is time-consuming, effort, and very uneconomical.
Cyprinid fish is the also maximum a group of the widest, kind that distributes in Cypriniformes, the total cyprinid fish 210 in the whole world belongs to, and more than 2000 kinds, is all freshwater fish, in the each large river of China, lake, extensively distribute, Ye Shi China is main cultivation fingerling in farmers' fish pond since ancient times.Therefore, the detection kit of exploitation cyprinid fish lipovitellin is with a wide range of applications and scientific value.
Summary of the invention
The object of the present invention is to provide a kind of kit and application thereof of qualitative detection cyprinid fish lipovitellin, to overcome the deficiencies in the prior art.
A kit for qualitative detection cyprinid fish lipovitellin, comprises a box body, in this box body, is equipped with: 1) 2 of pvdf membranes; 2) sheep anti mouse two of horseradish peroxidase-labeled resists 1; 3) each 1 of confining liquid, cleansing solution, nitrite ion, described cleansing solution is TBST; Confining liquid is the TBST containing 5% skimmed milk power; Nitrite ion is the 10mM Tris-HCl containing 0.06% (m/V) 3 '-diaminobenzidine; It is characterized in that this box body is also equipped with: 4) 1 of goldfish lipovitellin sterling; 5) 1 of mouse-anti goldfish lipovitellin polyclonal antibody.
Above-mentioned goldfish lipovitellin sterling is 100 μ g/ml goldfish lipovitellin solution.
The kit preparation method of above-mentioned qualitative detection cyprinid fish lipovitellin, comprises the following steps: 1) prepare goldfish lipovitellin sterling; 2) utilize step 1) the goldfish lipovitellin sterling that obtains prepares mouse-anti goldfish lipovitellin polyclonal antibody; 3) by step 1) 1 of the goldfish lipovitellin sterling, the step 2 that obtain) 1 of the mouse-anti goldfish lipovitellin polyclonal antiserum, 2 of the pvdf membranes that obtain, and anti-1 of the sheep anti mouse two of confining liquid, cleansing solution, nitrite ion and horseradish peroxidase-labeled packs box body jointly into, obtain the kit of cyprinid fish lipovitellin qualitative detection.
The application of mentioned reagent box in qualitative detection cyprinid fish lipovitellin.
The characteristic that qualification fish lipovitellin adopts it to contain sugar, phosphorus, fat group mostly both at home and abroad at present, adopt specific stain, amino acid composition analysis or analyze whether the albumen that its sugar, phosphorus, fat content carrys out purification Identification is lipovitellin, this method is time-consuming, effort, and very uneconomical.In addition amino acid composition analysis and sugar, phosphorus, fat assay are had relatively high expectations to instrument, complicated operation.And kit of the present invention utilizes the specific binding capacity between antigen-antibody, can be sensitive, the lipovitellin in qualitative detection cyprinid fish (comprising goldfish, carp, crucian, zebra fish etc.) ovary or prelarva body easily, and detectivity will be apparently higher than above method.The lipovitellin of researching fish has very big help to understanding fish early development situation, and the present invention can provide important references for growth and the reproductive status of evaluating fish.
Brief description of the drawings
Fig. 1 is the testing result of goldfish lipovitellin polyclonal antibody of the present invention to goldfish lipovitellin;
Fig. 2 is the testing result of goldfish lipovitellin polyclonal antibody of the present invention to Yolk Proteins In Cyprinus Carpio fat phosphoprotein;
Fig. 3 is the testing result of goldfish lipovitellin polyclonal antibody of the present invention to crucian lipovitellin;
Fig. 4 is the testing result of goldfish lipovitellin polyclonal antibody of the present invention to zebra fish lipovitellin.
Embodiment:
A kit for qualitative detection cyprinid fish lipovitellin, comprises a box body, in this box body, is equipped with: 1) 2 of pvdf membranes; 2) sheep anti mouse two of horseradish peroxidase-labeled resists 1; 3) each 1 of confining liquid, cleansing solution, nitrite ion, described cleansing solution is TBST; Confining liquid is the TBST containing 5% skimmed milk power; Nitrite ion is the 10mM Tris-HCl containing 0.06% (m/V) 3 '-diaminobenzidine; It is characterized in that this box body is also equipped with: 4) 1 of goldfish lipovitellin sterling; 5) 1 of mouse-anti goldfish lipovitellin polyclonal antibody.
Above-mentioned goldfish lipovitellin sterling is 100 μ g/ml goldfish lipovitellin solution.
In mentioned reagent box 4) goldfish lipovitellin sterling, prepare with following method:
Get the female goldfish ovary tissue that yolk forms the later stage, remove connective tissue, collect fish-egg, (25mM Tris-HCl, includes 70mM NaCl to add the homogenate buffer of 4 DEG C of precoolings of 3 times of volumes, 10mM EDTA and 1mM PMSF, pH7.5) mix, under condition of ice bath, use glass homogenizer homogenate, 4 DEG C, centrifugal 20 minutes of 10000g, collects supernatant; In supernatant, slowly add (NH
4)
2sO
4the saturation degree of powder to 70%, the liquid of saltouing; After 10 hours, at 4 DEG C, with 8000g centrifugal 10 minutes, supernatant discarded, was dissolved in 25mM Tris-HCl (including 0.07M NaCl, pH7.5) again by precipitation, obtained ovum homogenate extract;
Get 1ml ovum homogenate extract and add Sephacryl S-300 chromatographic column, with 25mM Tris-HCl (pH 7.5) wash-out containing 0.07M NaCl, collect the sample that contains goldfish lipovitellin for ion-exchange chromatography
(DEAE-Sepharose Fast flow), with the Tris-HCl damping fluid that contains respectively 0.07M, 0.1M, 0.2M, 0.3M and 1.0M NaCl
(25mM, pH 7.5)carry out discontinuous wash-out, collect the de-component of 0.2M, be accredited as goldfish lipovitellin sterling ,-80 DEG C of preservations.
In mentioned reagent box 5) mouse-anti goldfish lipovitellin polyclonal antibody, prepare with following method:
Adopt the mode of lumbar injection male mice, the goldfish lipovitellin 50 μ g of every injected in mice purifying, fully mix equal-volume antigen and Freund's complete adjuvant before injection; After this, the injection respectively at the 14th, 21,28,35,38 days, injected dose is 50 μ g, before injection, equal-volume antigen and Freund's complete adjuvant is fully mixed; Blood sampling in the 40th day, blood sampling is stopped eating the previous day.Adopt eye socket blood taking method to get blood, obtain antiserum for the preparation of mouse-anti goldfish lipovitellin polyclonal antibody;
Adopt the method for ammonium sulfate precipitation to prepare mouse-anti goldfish lipovitellin polyclonal antibody: first Mouse Antisera is mixed with negative control blood plasma 2:1,4 DEG C are rocked and spend the night, and the nonspecific antibody of centrifugal removal, then adds (NH
4)
2sO
4the saturation degree of powder to 20%, 4 DEG C are rocked 2 hours, centrifugal except defibrinating; Add 50% saturated ammonium sulfate, 4 DEG C are rocked 2 hours, the centrifugal albumin of removing; Add 33% saturated ammonium sulfate, 4 DEG C are rocked 2 hours; 4 DEG C, centrifugal 10 minutes of 8000g, obtains mouse-anti goldfish lipovitellin polyclonal antibody ,-80 DEG C of preservations.
Through above operation, the kit of the qualitative detection cyprinid fish lipovitellin obtaining is specifically composed as follows:
1) 2 of pvdf membranes;
2) 1 of goldfish lipovitellin sterling, is diluted to 1 μ g/ml with PBS before using;
3) 1 of mouse-anti goldfish lipovitellin polyclonal antibody, the volume ratio dilution of pressing 1:300 before using with confining liquid;
4) sheep anti mouse two of horseradish peroxidase-labeled resists 1, presses the volume ratio dilution of 1:500 before using with confining liquid;
5) each 1 of confining liquid, cleansing solution, nitrite ion, described cleansing solution is TBST (100mM Tris-HCl, 150mMNaCl, 0.05%Tween-20, pH7.5); Confining liquid is the TBST containing 5% skimmed milk power; Nitrite ion is the 10mM Tris-HCl containing 0.06% (m/V) 3 '-diaminobenzidine (DAB), uses in forward direction nitrite ion and adds 0.05% (v/v) hydrogen peroxide.
Kit of the present invention can be used for the qualitative detection of cyprinid fish lipovitellin.
The method of utilizing kit qualitative detection cyprinid fish lipovitellin of the present invention, specifically comprises the following steps:
1) will after the goldfish lipovitellin sterling in kit and Sample Dilution to be measured, carry out denaturing polyacrylamide gel electrophoresis, described sample to be measured comprises the homogenate of the albumen of purifying from fish ovary, female fish ovaries homogenate or prelarva;
2) albumen on electric current gel is transferred to pvdf membrane;
3) with the nonspecific binding site of confining liquid sealing pvdf membrane, 4 DEG C of overnight incubation, discard confining liquid;
4) add the mouse-anti goldfish lipovitellin polyclonal antibody (1:300) with confining liquid dilution, under room temperature, concussion is hatched, and discards solution, with cleansing solution washing pvdf membrane 3 times;
5) add the sheep anti mouse two anti-(1:500) by the horseradish peroxidase-labeled of confining liquid dilution, under room temperature, concussion is hatched, and discards solution, with cleansing solution washing pvdf membrane 3 times;
6) add nitrite ion, after protein band is clear, discard nitrite ion, with the reaction of distilled water color development stopping, pvdf membrane is preserved in lucifuge place after taking pictures;
7) in sample, find that there is colour developing band, the albumen that shows purifying is fish lipovitellin, and the depth of color and the width of band have represented that in sample, the content of lipovitellin is how many; In addition can utilize, the content of lipovitellin in the each sample of spectrodensitometry software semi-quantitative analysis.
As shown in accompanying drawing 1-4, its result is followed successively by the detection of Fig. 1 to goldfish lipovitellin; The detection of Fig. 2 to Yolk Proteins In Cyprinus Carpio fat phosphoprotein; The detection of Fig. 3 to crucian lipovitellin; The detection of Fig. 4 to zebra fish lipovitellin.Can see and on pvdf membrane, all occur obvious band, kit of the present invention and detection method thereof are having good application aspect qualitative detection cyprinid fish lipovitellin.
Claims (6)
1. a kit for qualitative detection cyprinid fish lipovitellin, comprises a box body, in this box body, is equipped with: 1) 2 of pvdf membranes; 2) sheep anti mouse two of horseradish peroxidase-labeled resists 1; 3) each 1 of confining liquid, cleansing solution, nitrite ion, described cleansing solution is TBST; Confining liquid is the TBST containing 5% skimmed milk power; Nitrite ion is containing 3 of mass volume ratio 0.06%, the 10mMTris-HCl of 3-diaminobenzidine; It is characterized in that this box body is also equipped with: 4) 1 of goldfish lipovitellin sterling; 5) 1 of mouse-anti goldfish lipovitellin polyclonal antibody.
2. kit according to claim 1, is characterized in that described goldfish lipovitellin sterling is 100 μ g/ml goldfish lipovitellin solution.
3. the kit of qualitative detection cyprinid fish lipovitellin as claimed in claim 1, is characterized in that the preparation method of described goldfish lipovitellin sterling is as follows:
Get the female goldfish ovary tissue that yolk forms the later stage, remove connective tissue, collect fish-egg, add the homogenate buffer of 4 DEG C of precoolings of 3 times of volumes to mix, use glass homogenizer homogenate under condition of ice bath, 4 DEG C, centrifugal 20 minutes of 10000g, collects supernatant; In supernatant, slowly add (NH
4)
2sO
4the saturation degree of powder to 70%, saltouts and spends the night; After 10 hours, at 4 DEG C, with 8000g centrifugal 10 minutes, supernatant discarded, was dissolved in 25mM Tris-HCl again by precipitation, includes 0.07M NaCl, and pH7.5 obtains ovum homogenate extract;
Get 1ml ovum homogenate extract and add Sephacryl S-300 chromatographic column, with the 25mM Tris-HCl containing 0.07M NaCl, pH7.5 wash-out; The sample that collection contains goldfish lipovitellin carries out ion-exchange chromatography, with the Tris-HCl damping fluid that contains respectively 0.07M, 0.1M, 0.2M, 0.3M and 1.0M NaCl, 25mM, pH7.5 carries out discontinuous wash-out, collect 0.2M elution fraction, be accredited as goldfish lipovitellin sterling.
4. the kit of qualitative detection cyprinid fish lipovitellin as claimed in claim 3, is characterized in that the preparation method of described mouse-anti goldfish lipovitellin polyclonal antibody is as follows:
Adopt the mode of lumbar injection male mice, the goldfish lipovitellin 50 μ g of every injected in mice purifying, fully mix equal-volume antigen and Freund's complete adjuvant before injection; After this, the injection respectively at the 14th, 21,28,35,38 days, injected dose is 50 μ g, before injection, equal-volume antigen and incomplete Freund's adjuvant is fully mixed; Blood sampling in the 40th day, blood sampling is stopped eating the previous day; Adopt eye socket blood taking method to get blood, obtain antiserum for the preparation of mouse-anti goldfish lipovitellin polyclonal antibody;
Adopt the method for ammonium sulfate precipitation to prepare mouse-anti goldfish lipovitellin polyclonal antibody: first Mouse Antisera is mixed with negative control blood plasma 2:1,4 DEG C are rocked and spend the night, and the nonspecific antibody of centrifugal removal, then adds (NH
4)
2sO
4the saturation degree of powder to 20%, 4 DEG C are rocked 2 hours, centrifugal except defibrinating; Add 50% saturated ammonium sulfate, 4 DEG C are rocked 2 hours, the centrifugal albumin of removing; Add 33% saturated ammonium sulfate, 4 DEG C are rocked 2 hours; 4 DEG C, centrifugal 10 minutes of 8000g, obtains mouse-anti goldfish lipovitellin polyclonal antibody.
5. cyprinid fish lipovitellin qualitative detection kit claimed in claim 1, is characterized in that the application of this kit in the qualification of cyprinid fish lipovitellin.
6. the method for utilizing the kit qualitative detection cyprinid fish lipovitellin described in claim 1, is characterized in that the method comprises the following steps:
1) will after the goldfish lipovitellin sterling in kit and Sample Dilution to be measured, carry out denaturing polyacrylamide gel electrophoresis, described sample to be measured comprises the homogenate of the albumen of purifying from fish ovary, female fish ovaries homogenate or prelarva;
2) albumen on running gel is transferred to pvdf membrane;
3) with the nonspecific binding site of confining liquid sealing pvdf membrane, 4 DEG C of overnight incubation, discard confining liquid;
4) add with confining liquid by the mouse-anti goldfish lipovitellin polyclonal antibody of the volume ratio dilution of 1:300, under room temperature, concussion is hatched 4 hours, discards solution, with cleansing solution washing pvdf membrane 3 times;
5) add with confining liquid anti-ly by the sheep anti mouse of the horseradish peroxidase-labeled of the volume ratio dilution of 1:500 two, under room temperature, concussion is hatched 4 hours, discards solution, with cleansing solution washing pvdf membrane 3 times;
6) add nitrite ion, after protein band is clear, discard nitrite ion, with the reaction of distilled water color development stopping, pvdf membrane is preserved in lucifuge place after taking pictures;
7) in sample, find that there is colour developing band, the albumen that shows purifying is fish lipovitellin, and the depth of color and the width of band have represented the content of lipovitellin in sample; Or utilize the content of lipovitellin in the each sample of spectrodensitometry software semi-quantitative analysis.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201210557080.1A CN103033630B (en) | 2012-12-20 | 2012-12-20 | Kit for qualitative detection of lipovitellin of cyprinid fish and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201210557080.1A CN103033630B (en) | 2012-12-20 | 2012-12-20 | Kit for qualitative detection of lipovitellin of cyprinid fish and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN103033630A CN103033630A (en) | 2013-04-10 |
| CN103033630B true CN103033630B (en) | 2014-10-22 |
Family
ID=48020738
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201210557080.1A Expired - Fee Related CN103033630B (en) | 2012-12-20 | 2012-12-20 | Kit for qualitative detection of lipovitellin of cyprinid fish and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN103033630B (en) |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102288754A (en) * | 2010-10-09 | 2011-12-21 | 中国疾病预防控制中心寄生虫病预防控制所 | Kit adopting specific polyclonal antibody for detection of agkistrodon acutus tongue-shaped worm circulating antigen |
| CN102768282A (en) * | 2012-07-20 | 2012-11-07 | 湖北省农业科学院农产品加工与核农技术研究所 | Preparation and detection methods of kit for detecting chub vitellogenin (VTG) |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20090274660A1 (en) * | 1999-08-17 | 2009-11-05 | Immunopath Profile, Inc. | Pluripotent therapeutic compositions and uses thereof |
-
2012
- 2012-12-20 CN CN201210557080.1A patent/CN103033630B/en not_active Expired - Fee Related
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102288754A (en) * | 2010-10-09 | 2011-12-21 | 中国疾病预防控制中心寄生虫病预防控制所 | Kit adopting specific polyclonal antibody for detection of agkistrodon acutus tongue-shaped worm circulating antigen |
| CN102768282A (en) * | 2012-07-20 | 2012-11-07 | 湖北省农业科学院农产品加工与核农技术研究所 | Preparation and detection methods of kit for detecting chub vitellogenin (VTG) |
Non-Patent Citations (9)
| Title |
|---|
| 三种方法纯化相思子毒素单抗的比较;罗胜军等;《动物医学进展》;20061231;第27卷(第11期);72-75 * |
| 师晓栋.玫瑰无须鲃卵黄蛋白原和卵黄脂磷蛋白的纯化、鉴定、免疫分析和功能研究.《中国海洋大学研究生学位论文》.2004,正文第54-58页. |
| 改良的硫酸铵沉淀法纯化大雁的卵黄抗体;杨帆等;《中国比较医学杂志》;20071130;第17卷(第11期);625-628 * |
| 杨帆等.改良的硫酸铵沉淀法纯化大雁的卵黄抗体.《中国比较医学杂志》.2007,第17卷(第11期),625-628. |
| 玫瑰无须鲃卵黄蛋白原和卵黄脂磷蛋白的纯化、鉴定、免疫分析和功能研究;师晓栋;《中国海洋大学研究生学位论文》;20041231;正文第54-58页 * |
| 罗胜军等.三种方法纯化相思子毒素单抗的比较.《动物医学进展》.2006,第27卷(第11期),72-75. |
| 邴欣.金鱼卵黄原蛋白多克隆抗血清制备及在内分泌扰乱化学物质检测中的应用.《工程科技辑》.2007,全文. |
| 邴欣等.真鲷（Pagrosomus major）卵黄原蛋白的分离纯化及抗血清制备.《高技术通讯》.2010,第20卷(第12期),1309-1313. * |
| 金鱼卵黄原蛋白多克隆抗血清制备及在内分泌扰乱化学物质检测中的应用;邴欣;《工程科技辑》;20070215;全文 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN103033630A (en) | 2013-04-10 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Manahan et al. | Bacteria-free sea urchin larvae: selective uptake of neutral amino acids from seawater | |
| Whyte et al. | Biochemical and energy changes during embryogenesis in the rock scallop Crassadoma gigantea | |
| Finn et al. | Respiration and nitrogen metabolism of Atlantic halibut eggs (Hippoglossus hippoglossus) | |
| Beckman et al. | Egg production by Acartia tonsa in Long Island sound | |
| CN105899950B (en) | In fish scale as the biomarker for chronic stress glucocorticoid quantization | |
| Fischer | ELEMENTS OF ENERGY BALANCE IN GRASS CARP(CTENOPHARYNGODON IDELLA VAL.): III. ASSIMILABILITY OF PROTEINS, CARBOHYDRATES, AND LIPIDS BY FISH FED WITH PLANT AND ANIMAL FOOD | |
| Quin | 2-Aminoethylphosphonic acid in insoluble protein of the sea anemone Metridium dianthus | |
| Nynca et al. | Shotgun proteomics of rainbow trout ovarian fluid | |
| El-Hanoun et al. | Effect of magnetized well water on blood components, immune indices and semen quality of Egyptian male geese. | |
| Cejko et al. | Sperm quality and selected biochemical markers of seminal plasma at the beginning of the reproductive period of common carp, Cyprinus carpio L. | |
| Pacheco et al. | Identification of the Toxic Pentapeptide Nodularin in a Cyanobacterial Bloom in a Shrimp Farm in South American Atlantic Coast. Pharm Anal Acta 7: 479. doi: 10.4172/2153-2435.1000479 Page 2 of 3 Pharm Anal Acta ISSN: 2153-2435 PAA, an open access journal Volume 7• Issue 5• 1000479 Table 1: Productivity of the white shrimp Litopenaeus vannamei in the tanks with, and without, Nodularia (NOD) blooms | |
| CN103033630B (en) | Kit for qualitative detection of lipovitellin of cyprinid fish and application thereof | |
| CN104569423B (en) | Zebra fish vitellogenin indirect competitive ELISA test kit and detection method thereof and application | |
| Gilles | Osmoregulation in the Stenohaline Crab «Libinia Emarginata» Leech | |
| Comeau et al. | Key Arctic pelagic mollusc (Limacina helicina) threatened by ocean acidification. | |
| CN104597247B (en) | Zebra fish vitellogenin sandwich ELISA lcits and detection method thereof and application | |
| Abd El-Salaam et al. | Thermoregulation and blood biochemical changes in male dromedary camels during hot-humid and hot-dry environments under Egyptian conditions | |
| CN104360075B (en) | Greenling vitellogenin indirect ELISA reagent kit and preparation method thereof, detection method and application | |
| Fang et al. | Tolerance, oxygen consumption and ammonia excretion of Ophiopholis sarsii vadicola in different temperatures and salinities | |
| CN103910798A (en) | Antibody for resisting vipera berus toxin and long-noded pit viper toxin and preparation method and application thereof | |
| CN104569384B (en) | Zebra fish vitellogenin HP immunoblotting kit and detection method thereof and application | |
| Thyrring et al. | Ocean acidification increases susceptibility to sub-zero air temperatures in ecosystem engineers and limits poleward range shifts | |
| CN107367617A (en) | A kind of kit of the quantitative detection seawater medaka vitellogenin of improvement, preparation method and application | |
| Al-Toum et al. | Water deprivation and its effect on some blood constituents in Idmi Gazelle, Gazella gazella | |
| Samaee et al. | Quantitative composition of vitellogenin‐derived yolk proteins and their effects on viability of embryos and larvae of common dentex (Dentex dentex), a marine pelagophil teleost |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20141022 Termination date: 20151220 |
|
| EXPY | Termination of patent right or utility model |