CN102174367A - Artificial rumen device used for continuous multiple determination of pH value - Google Patents
Artificial rumen device used for continuous multiple determination of pH value Download PDFInfo
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- CN102174367A CN102174367A CN2011100067845A CN201110006784A CN102174367A CN 102174367 A CN102174367 A CN 102174367A CN 2011100067845 A CN2011100067845 A CN 2011100067845A CN 201110006784 A CN201110006784 A CN 201110006784A CN 102174367 A CN102174367 A CN 102174367A
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- sealing plug
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M23/00—Constructional details, e.g. recesses, hinges
- C12M23/46—Means for fastening
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M33/00—Means for introduction, transport, positioning, extraction, harvesting, peeling or sampling of biological material in or from the apparatus
- C12M33/04—Means for introduction, transport, positioning, extraction, harvesting, peeling or sampling of biological material in or from the apparatus by injection or suction, e.g. using pipettes, syringes, needles
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
- C12M41/26—Means for regulation, monitoring, measurement or control, e.g. flow regulation of pH
Abstract
The invention discloses an artificial rumen device used for continuous multiple determination of a pH value, and relates to the technical field of animal husbandry scientific research appliances. The device comprises a gas generation syringe, and a fermentation device connected with an injection end of the gas generation syringe. In the invention, a sealed container-shaped fermentation device only with a round opening is designed; the opening is provided with a sealing plug; the sealing plug impacts and seals the round opening; the sealing plug is provided with a gas-guide tube penetrating through the center of the sealing plug; one end of a silicone tube is connected with the gas-guide tube; the other end of the silicone tube is connected with the injection end of the gas generation syringe; the test determination result and the traditional device have no obvious difference and good consistency, thus the influence on the fermentation result caused by the factors, such as the gravity and the air leakage and the like of the gas generation syringe is overcome, and the device is convenient in use, replaces the traditional gas generation cylinder device to determine the pH value of rumen liquid, and realizes single-sample continuous multiple sampling to determine the pH value; the time point needs not to be used as a processing factor, the sample size is greatly shortened, and the manual amount and the implementation period are obviously lowered.
Description
Technical field
The invention belongs to livestock industry scientific research utensil technical field, relate to a kind of artificial rumen device of simulating the ruminant tumor gastric fermentation.
Background technology
The aerogenesis syringe is the artificial rumen device of in-vitro simulated ruminant tumor gastric fermentation in the herding industry, uses more general.Usually, the pH value that needs continuous a plurality of point in time sampling to measure rumen fluid reflects the pH dynamic change of rumen fluid, but, be subjected to the restriction on aerogenesis syringe device shape and the Capacity design, every aerogenesis syringe can only be got a sample and measure the pH value, and each sample then needs and will count the time as the processing factor, causes sample size bigger, waste time and energy, and increased the risk and the implementation cost of systematic error.Aerogenesis syringe simulation rumen fermentation unit is a kind of erect type device simultaneously, external aerogenesis result will be subjected to the influence of piston gravity to the result, and to seal with tongs in the syringe lower end, might leak gas or intake and influence experimental result, it is easily broken to add the aerogenesis syringe, makes the experimental implementation inconvenience.We have carried out new design at the support energetically to this research under national beef cattle industries technical system.Literature search finds, with the immediate patent No. of the present invention be the patent document of 01104466.7 " a kind of two outer flow pattern laboratory ferment jars of simulation rumen fermentation " by name.This patent disclosure a kind of two outer flow pattern laboratory ferment jars that are used to simulate ruminant tumor gastric fermentations such as cattle and sheep.Technical scheme comprises tank body, agitator, motor, heating unit, charging opening, oxygenless gas import, artificial saliva skirt, solid phase outflow pipe, feature is, the sidewall of tank body be provided with one outwards outstanding and communicate, be equipped with in it tubulose emergence of filter screen with tank interior; Heating unit is a water-bath.It is even to have the fermentation contents stirred, the unanimity of being heated, and simulate effect is good, advantages such as liquid phase mesh screen disassembling, assembling and replacing convenience.But purpose of the present invention, technical scheme and technical characterictic and described patent are different.
Summary of the invention
The objective of the invention is to overcome the shortcoming of above-mentioned traditional aerogenesis needle cylinder device, provide a kind of fermentation volume big and easy to use, realize each sample energy continuous several times sampling determination pH value, significantly reduce sample size, overcome of the influence of piston gravity, reduce the rumen zymosis device of systematic error risk detected result.The technical solution that realizes the present invention's purpose is such.
But a kind of continuous several times is measured the artificial rumen device of pH value, comprise aerogenesis syringe and a kind of fermentation unit that links to each other with its dispensing end, fermentation unit is a sealed vessel, one circular open is only arranged, sealing plug is arranged on the opening, and sealing plug is that caulked seals up circular open, and a conduit that penetrates the sealing plug center of circle is arranged on the sealing plug, have an end of a silicone tube to be connected with described conduit, the other end of silicone tube links to each other with the dispensing end of described air tube tube.
The present invention compares with traditional aerogenesis cartridge, has following beneficial effect:
(1) device provided by the invention substitutes traditional aerogenesis cartridge and measures the rumen fluid ph value, and its measurement result and conventional apparatus do not have significant difference, high conformity;
(2) this device is realized single sample continuous several times sampling determination pH value, does not then need time point significantly to dwindle sample size as the processing factor, significantly reduces and manually measures and the implementation cycle;
(3) each step number of operations of mensuration is greatly reduced, all ingredients consumption reduces, thereby reduces systematic error and implementation cost;
(4) overcome because of of the influence of factors such as aerogenesis cylinder piston gravity and gas leakage the fermentation result, easy to use.
Description of drawings
Fig. 1 is an artificial rumen apparatus structure synoptic diagram of the present invention.
Fig. 2 is traditional aerogenesis needle cylinder device structural representation.
Fig. 3 is that the pH of traditional aerogenesis syringe and artificial rumen device measures curve.
Symbolic representation among the figure: 1, aerogenesis syringe; 1.2, dispensing end; 1.3, tongs; 2, fermentation unit; 2.1, sealing plug, 2.2, airway, 3, silicone tube.
Embodiment
Referring to Fig. 1 Fig. 2 as can be known, traditional aerogenesis syringe 1 and silicone tube 3 and tongs 1.3 of linking to each other with its dispensing end 1.2 of comprising.The present invention has improved the structure of traditional aerogenesis tube, design a kind of fermentation unit 2, fermentation unit 2 is sealed vessels that a circular open is only arranged, sealing plug 2.1 is arranged on the opening, sealing plug 2.1 is that caulked seals up circular open, one airway 2.2 that penetrates sealing plug 2.1 centers of circle is arranged on the sealing plug 2.1, have an end of a silicone tube 3 to be connected with described conduit 2.2, the other end of silicone tube 3 links to each other with the dispensing end 1.2 of described aerogenesis syringe 1.
What present embodiment fermentation unit 2 adopted is that the glass triangle bottle is made fermentation unit, 9 centimetres of its base diameters, 3 centimetres of bottleneck diameters; The opening of fermentation unit 2 has the sealing plug 2.1 of tight plug bottleneck, and what sealing plug 2.1 was selected for use is soft rubber ball; One airway 2.2 that penetrates the soft rubber ball center is arranged on the soft rubber ball, and what airway 2.2 adopted is Glass tubing; There is an end of a silicone tube 3 to be connected with described airway 2.2, the other end of silicone tube 3 links to each other with the dispensing end 1.2 of described aerogenesis syringe 1, silicone tube 3 length are changeable, decide on operation, present embodiment is selected length for use is 5 centimetres, has formed the artificial rumen device of can continuous several times measuring the pH value.This device by following evidence apparatus of the present invention in theoretical or practice, all be feasible.
The following example of the present invention is the pH pH-value determination pH test comparing result of device provided by the invention and traditional aerogenesis needle cylinder device.
Experimental example:
Select 3 healthy fistula beef cattles as for the examination animal, raise the rumen fluid of gathering in back 2 hours morning and filter, configure artificial rumen liquid and substrate according to the appendix method through four layers of antiseptic gauze.Reference examples (aerogenesis tube group), experimental example (fermentation flask group) are established in experiment, and 6 time points are established in 5 repetitions of the every time point of precedent altogether, totally 30 samples, and back example is 5 samples of totally 5 repetitions.The substrate component of reference examples, experimental example is identical, and (corn: fresh grain stillage=5: 5), each substrate sample amount of fermentation of back example is the former 8 times.Concrete consumption sees the following form:
Table 1 is respectively organized artificial rumen liquid and substrate consumption
Experiment yeast phase 36h, respectively the 3rd, 8,14,18,24, measure the pH value during 36h.Two treatment group are operated according to following steps:
One, material is prepared:
The preparation of feed substrate: air-dry corn was pulverized 40 mesh sieves, and extracting screen underflow is standby, and fresh white vinasse oven dry back appropriateness is crushed to granularity and can gets final product by the fermentation flask conduit.
Aerogenesis syringe: choose 30 100ml glass syringes, note checking the uniformity coefficient of piston resistance to air loss and piston resistance, 30 of syringe needle place cover plug thin silicone tube (the about 0.4cm of internal diameter is about 6cm, 30) tongses.
Fermentation flask: choose 5 250ml glass triangle bottles, 5 are turned over the cap soft rubber ball, with the about 0.4cm of internal diameter, the short Glass tubing that is about 7cm passes soft rubber ball, and the lower end exceeds the about 1.5cm of soft rubber ball, behind the cover plug 0.6cm internal diameter silicone tube of upper end, with wearing the soft rubber ball and the triangular flask jam-pack of conduit, be used to collect gas with the 100ml syringe jacket at the sebific duct the other end, infusion bottle and catheter interface all place water, slowly push each junction of gas checking device and have or not bubble to produce, no bubble is emerged and can be used.Thick 5 of silicone tubes (the about 0.6cm of internal diameter is about 6cm), 5 of tongses, above-mentioned article are cleaned the oven dry back and are used.
Other equipment: but 2 in temperature adjustment shaking bath pot, 1 in baking oven, 1 of electronic balance, 1 of pH meter.
Two, place substrate:
The fermentation substrate that makes places cleaning exsiccant 100ml aerogenesis syringe and 250ml glass triangle bottle respectively according to last table consumption.
The aerogenesis syringe: Vaseline is evenly smeared at preceding 1/3 position of piston, and slowly push piston is got rid of most of air, and headgear plug silicone tube is clamped sebific duct with tongs;
Fermentation flask: slowly feed carbon dioxide, cover soft rubber ball rapidly, clamp sebific duct.
Three, inject rumen fluid:
The aerogenesis syringe: open tongs, after injecting by sebific duct with disposable aerogenesis syringe absorption 30ml artificial rumen liquid, aerogenesis syringe push piston is up discharged unnecessary bubble, clamps tongs rapidly.
Fermentation flask: take out soft rubber ball, rapidly 240ml artificial rumen liquid is injected bottle, cover soft rubber ball, clamp sebific duct with tongs immediately and keep airtight.
Four, water-bath is cultivated:
Aerogenesis syringe: stand upside down and place on the shaking bath.
Fermentation flask: go to the water-bath shaking table, take off tongs, the 100ml glass aerogenesis syringe that the sebific duct other end is inserted begins concussion and cultivates to receive the gas (aerogenesis tube) that fermentation produces.
Bath temperature is set 39 ℃, is as the criterion with water temperature in the same mensuration of the thermometer accurately shaking table.60 rev/mins of oscillation frequency.
Five, pH pH-value determination pH:
At the setting-up time point, measure as follows respectively.
The aerogenesis syringe: 5 that take out respectively rapidly this time point are repeated samples, take off tongs after shaking all, push 3ml liquid in the 15ml centrifuge tube, go to pH meter, and probe is inserted mensuration liquid level under.This batch injection tube stops fermentation after the sampling, does not re-use.
Fermentation flask: gas in the aerogenesis tube is pushed about 3ml to fermentation flask, from water-bath, take out vial rapidly and shake equal, sebific duct inserts clean 10ml syringe, be inverted fermentation flask rapidly and extract the 3ml mixed solution and go to the 15ml centrifuge tube with the aerogenesis syringe, take back sebific duct rapidly after the exhaust of aerogenesis tube, fermentation flask is put back to water-bath and is continued fermentation.The pH meter probe inserts the centrifuge tube liquid level and measures the pH value down.
Six, measurement result (referring to Fig. 3):
The pH mean value of table 2 liang each time point of group
Table 3SPSS 12.0 ANOVA (variance analysis) result
Draw two kinds of pH value differences different not remarkable (P=0.824) that method records by table 3.
Table 4SPSS12.0 correlation test result:
Two groups of pH value curves are seen Fig. 3.Table 4 shows, two the two poles of the earth significant correlations between the pH curve of two kinds of device mensuration.(P＝0.998)
The above results shows that device provided by the present invention is than significantly not outstanding difference of the pH pH-value determination pH result of conventional apparatus and height correlation, and alternative conventional apparatus is fully feasible.
Below derivation advantage of the present invention be, significantly reduced sample number, manually measured and the enforcement time.
Tradition aerogenesis cartridge limits because of itself, need be with time point as the processing factor, and its total sample size formula is:
T＝t×C×r ①
And fermentation flask device of the present invention has overcome the restriction of conventional apparatus, realizes the continuous multi-site determination of each sample, then need not time point as the processing factor, and its total sample size formula is:
T＝C×r ②
T: the set time counts, C: sample process number, r: the repeat number of each sample.
By formula 1., 2., to establish 5 time points usually, repeat number is 5, and treatment group is 4, and yeast phase 24h is an example, and two kinds of scheme contrasts are as shown in the table:
The comparison of two kinds of devices of table 5
Annotate: only count the actually operating time when artificial; Implementation cycle contains setup time by working days, all estimates by empirical value
By table 5 as seen, device provided by the invention is the reduced sample amount greatly, significantly reduces artificial amount and experimental period, implements efficient thereby improve.Simultaneously, reduce systematic error and implementation cost.
Appendix:
Artificial rumen liquid making method (good together with reference to cloth, 2006 method):
Rumen fluid is mixed by 1: 2 volume ratio with artificial saliva, and stirring gets final product.Artificial saliva is prepared as follows.
520.2 ml distilled water+0.1 milliliter trace element solution (A)+208.1ml damping fluid (B)+208.1ml
Major element solution (C)+1.0ml resazurin solution (D), with carbon dioxide saturated and be warming up to 39 ℃ after, add 62.4ml reduced liquid (E), continue to feed CO
2, until solution by light blue change into colourless.
A, B, C, D, each solution formula of E of being modulated into artificial saliva are as follows:
A solution: 13.2gCaCl
2H
2O+10.0gMnCl
24H
2O+1.0gCoCl
26H
2O+8gFeCl
36H
2O, the adding distil water dissolving is settled to 1000ml;
B solution: 4.0gNH4HCO
3+ 35gNaHCO
3, the adding distil water dissolving is settled to 1000ml;
C solution: 5.7gNa
2HPO
4(anhydrous)+6.29KH
2PO
4(anhydrous)+0.6gMgSO
47H
2O, the adding distil water dissolving is settled to 1000ml;
D solution: the 100mg resazurin is dissolved in 100ml distilled water;
E solution: 4.0ml NaOH (1N)+625mgNa
2S9H
2O+95ml distilled water.
Claims (1)
1. but a continuous several times is measured the artificial rumen device of pH value, comprise, aerogenesis syringe (1) and a kind of fermentation unit (2) that links to each other with its dispensing end (1.2), it is characterized in that: described fermentation unit (2) is the sealed vessel that a circular open is only arranged, and sealing plug (2.1) is arranged on the opening; Described sealing plug (2.1) is that caulked seals up circular open, one airway (2.2) that penetrates sealing plug (2.1) center of circle is arranged on the sealing plug (2.1), have an end of a silicone tube (3) to be connected with described airway (2.2), the other end of silicone tube (3) links to each other with the dispensing end (1.2) of described aerogenesis syringe (1).
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110499242A (en) * | 2019-09-24 | 2019-11-26 | 扬州大学 | A kind of simulator and its analogy method of cud epithelial tissue in vitro culture |
| CN114106989A (en) * | 2021-11-18 | 2022-03-01 | 黑龙江八一农垦大学 | Rumen in-vitro fermentation device and method thereof |
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-
2011
- 2011-01-13 CN CN2011100067845A patent/CN102174367A/en active Pending
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| CN2836723Y (en) * | 2005-08-26 | 2006-11-15 | 中国农业科学院畜牧研究所 | Device for real time and continuous monitoring ruminant rumen liquor pH value |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110499242A (en) * | 2019-09-24 | 2019-11-26 | 扬州大学 | A kind of simulator and its analogy method of cud epithelial tissue in vitro culture |
| CN114106989A (en) * | 2021-11-18 | 2022-03-01 | 黑龙江八一农垦大学 | Rumen in-vitro fermentation device and method thereof |
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Application publication date: 20110907 |