CN103168744B - Method for judging physical conditions of brown-eared pheasant chickens through non-invasive sampling method and for grouping feeding - Google Patents
Method for judging physical conditions of brown-eared pheasant chickens through non-invasive sampling method and for grouping feeding Download PDFInfo
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Abstract
The invention discloses a method for judging physical conditions of brown-eared pheasant chickens through a non-invasive sampling method and for grouping feeding, in particular to a brown-eared pheasant brooding method. According to the brown-eared pheasant brooding method, the brown-eared pheasant chickens to be bred are grouped and isolated for being fed according to the different contents of immune globulin A in excrement. Experiments prove that the healthy conditions of the brown-eared pheasant chickens can be evaluated through monitoring of the contents and the changes of the immune globulin A in the excrement of the brown-eared pheasant chickens, and the method is an ideal non-invasive method. When the method is used in the brood process of brown-eared pheasants, and the chickens are grouped and isolated for being fed according to the contents of the immune globulin A in the excrement of the chickens to be bred, results show that compared with comparison groups which are allocated in a random mode, the feeding method greatly lowers the death rate of the chickens for the whole chicken group.
Description
Technical field
The present invention relates to a kind of judge brown eared pheasant chick physical condition and raise in the grouping method of brooding, particularly a kind of brown eared pheasant method of brooding.
Background technology
Brown eared pheasant (Crossoptilon mantchuricum) is Chinese distinctive Precious, Rare, Endangered birds, animals under first-class state protection.Due to this species Habitat Fragmentation, population isolation; cause fertility degeneration, population decline; World Conservation Union (IUCN), international birds alliances (BirdLife), government utility development administration (WPA) are classified as global endangered species, are listed in annex I by endangered species of wild fauna and flora international trade pact (CITES).Detecting and assess the survival condition of rare and endangered species, and implement necessary manually managing and protecting, is to ensure to raise population health quality, put the key link of returning field long-term surviving and set up wild stocks, but current this respect research work is very few.Trace it to its cause, first, the pheasant class animal of free living is alert flexibly, and scope of activities is wide, and habitat is hidden, wants to obtain its tissue samples very difficult; Secondly, chick birth is small and weak, and seizure and blood sample collection product are very strong environmental stimulis, very easily cause chick Human Related Deaths; Moreover chick growing is to subadult, the operational difficulties of undamaged effectively long-term its healthy variation of monitoring is heavy, has had a strong impact on the work of brooding of brown eared pheasant.Obviously, be subject to the restriction of the technical elements such as sample collection and mensuration, do not have so far the physiological research of wild pheasant class Immune Profile In Chicks.In Chicks ' Growth process, immune physiological variation is the important channel of evaluating health status.Therefore,, as the physiological important indication index of immunity, the monitoring of immunoglobulin content and variation is particularly important.
The physiological situation monitoring of brown eared pheasant Immune Profile In Chicks globulin content, traditional animal bloodletting method cannot be suitable for, need a kind of simple, quick, the non-invasive method of development of exploring badly, for judging brown eared pheasant chick physical condition and raising to brood and hive off.
Summary of the invention
The object of this invention is to provide a kind of method of hiving off and brooding in brown eared pheasant chick physical condition and raising of judging.
The brown eared pheasant provided by the present invention method of brooding, for waiting to educate brown eared pheasant chick according to the isolated rearing of dividing into groups of the content difference of immunoglobulin A in ight soil.
In described method, described brown eared pheasant chick can be the brown eared pheasant of 2～14 ages in days.In one embodiment of the invention, described brown eared pheasant chick is specially the brown eared pheasant of 2～4 ages in days, 7～9 ages in days or 12～14 ages in days.
Further, the brown eared pheasant provided by the present invention method of brooding, can be any in following (1)～(3):
(1) 2～4 ages in days are waited to educate brown eared pheasant chick and are divided into following three groups, isolated rearing between group:
Group A: in ight soil, the content of immunoglobulin A is lower than waiting to educate brown eared pheasant chick described in 834ng/g;
Group B: in ight soil, the content of immunoglobulin A is higher than waiting to educate brown eared pheasant chick described in 1133ng/g;
Group C: in ight soil the content of immunoglobulin A between 834ng/g and 1133ng/g described in wait to educate brown eared pheasant chick;
(2) 7～9 ages in days are waited to educate brown eared pheasant chick and are divided into following three groups, isolated rearing between group:
Group A: in ight soil, the content of immunoglobulin A is lower than waiting to educate brown eared pheasant chick described in 1196ng/g;
Group B: in ight soil, the content of immunoglobulin A is higher than waiting to educate brown eared pheasant chick described in 1363ng/g;
Group C: in ight soil the content of immunoglobulin A between 1196ng/g and 1363ng/g described in wait to educate brown eared pheasant chick;
(3) 12～14 ages in days are waited to educate brown eared pheasant chick and are divided into following three groups, isolated rearing between group:
Group A: in ight soil, the content of immunoglobulin A is lower than waiting to educate brown eared pheasant chick described in 1298ng/g;
Group B: in ight soil, the content of immunoglobulin A is higher than waiting to educate brown eared pheasant chick described in 1556ng/g;
Group C: in ight soil the content of immunoglobulin A between 1298ng/g and 1556ng/g described in wait to educate brown eared pheasant chick.
In said method, waiting to educate brown eared pheasant chick described in described group of A in (1)～(3), to be health status not good, and dead chick very easily occurs; Described in described group of C, waiting to educate brown eared pheasant chick, to be health status good, is not easy to occur dead chick; Described in described group of B, waiting to educate brown eared pheasant chick is the chick of health status between group A and group C.
A further object of the present invention is to provide kit and the application thereof of a kind of qualification or the brown eared pheasant chick of assistant identification health status.
Application provided by the present invention is specially that material for quantitatively detecting Chicken immunoglobulin A is identified taking ight soil (fresh excreta) as sample to be tested in preparation or the application of the kit of the brown eared pheasant chick of assistant identification health status.
In described application, the described material for quantitative detection Chicken immunoglobulin A can be Chicken immunoglobulin A specific antibody or Chicken immunoglobulin A enzyme linked immunological kit.
In one embodiment of the invention, described is Chicken immunoglobulin A enzyme linked immunological kit for the material that quantitatively detects Chicken immunoglobulin A, be specially the Chicken immunoglobulin A enzyme linked immunological kit of Bethyl company of the U.S., its catalog number is E30-103, be Chicken IgA ELISA Quantitation Set(E30-103, Bethyl Laboratories, USA).
The kit of the brown eared pheasant chick of qualification provided by the present invention or assistant identification health status, comprises coated antibody, enzyme mark thing and fresh excreta collection kit; Described coated antibody is Chicken immunoglobulin A specific antibody, and described enzyme mark thing is Chicken immunoglobulin A enzyme labelled antibody.
Small-sized airtight plastic casing, glass receiving flask processed, valve bag etc. that described fresh excreta collection kit can be disposable glove, tweezers, can uncap.
Described kit also comprises as lower at least one: Chicken immunoglobulin A standard solution, Chicken immunoglobulin extract, substrate nitrite ion, stop buffer, cleaning solution, coated buffer solution and confining liquid.
In described kit, described Chicken immunoglobulin A standard solution can be the standard liquid of the multiple concentration within the scope of finite concentration, for example can be between 0～1000ng/mL, as concentration is respectively 1000.00ng/mL, 500.00ng/mL, 250.00ng/mL, 125.00ng/mL, 62.50ng/mL, 31.25ng/mL, 15.60ng/mL.
In described kit, described Chicken immunoglobulin extract is pH7.2, the phosphate buffer of the 0.01M that contains polysorbas20; The volumn concentration of described polysorbas20 in described Chicken immunoglobulin extract is 0.05%.The solvent of the phosphate buffer of described 0.01M is water, and solute and concentration thereof are as follows: NaCl8.50g/L, Na
2hPO
42.20g/L and KH
2pO
40.30g/L.
In described kit, described substrate nitrite ion, described stop buffer, described cleaning solution, described coated buffer solution and described confining liquid are the conventional ELISA reagent in this area.
In the use of described kit, the ight soil of brown eared pheasant chick to be measured (fresh excreta) is fully mixed according to the ratio of 1g:10mL with described Chicken immunoglobulin extract, obtain mixture; Mixture is centrifugal, get supernatant, use enzyme linked immunosorbent assay described supernatant to be carried out to the quantitative detection of immunoglobulin A, further calculate the content of immunoglobulin A in the ight soil of Unit Weight.
Another object of the present invention is to provide the preparation method of the above kit.
The preparation method of described kit, specifically can comprise the step of (1) or (2) as follows:
(1) step of described coated antibody, described enzyme mark thing and described fresh excreta collection kit being packed separately respectively;
(2) by described coated antibody, described enzyme mark thing, described fresh excreta collection kit with as lower at least one step of packaging separately respectively: as described in Chicken immunoglobulin A standard solution, as described in Chicken immunoglobulin extract, as described in substrate nitrite ion, as described in stop buffer, as described in cleaning solution, as described in coated buffer solution and as described in confining liquid.
All described chick all can be the brown eared pheasant of 1～33 age in days above; As 2～4 ages in days, 7～9 ages in days, 12～14 ages in days; For another example 3 ages in days, 8 ages in days, 13 ages in days, 18 ages in days or 33 ages in days.
All described brown eared pheasant chick health status, in the present invention, are all embodied in this index of lethality or survival rate above.
In the present invention, lethality, higher than 70%, is considered as health status poor; Lethality, lower than 35%, is considered as health status good.
Experimental results show that, content and the brown eared pheasant chick of Assessment of Changes health status by monitoring ight soil immunoglobulin A are desirable non-damage methods, applied to brooding in process of brown eared pheasant, according to chicken manure to be brooded just in the content of immunoglobulin A the chick isolated rearing of dividing into groups, result shows compared with the control group of random packet, for whole chick colony, this feeding manner greatly reduces the lethality of chick.The present invention is for protection and the artificial breeding meaning of crucial importance of the brown eared pheasant of the peculiar endangered species of China.
Brief description of the drawings
Fig. 1 is Chicken immunoglobulin A calibration curve.
Fig. 2 is the FA mean value of the brown eared pheasant chick of each age in days.
Fig. 3 is the dead individual ight soil IgA(DFA of brown eared pheasant chick) content and the individual ight soil IgA(SFA of surviving) the vertical scatter diagram of content.
Fig. 4 is the block diagram of brown eared pheasant chick DFA content and SFA content, and wherein * is significant difference (P < 0.05), and * * is utmost point significant difference (P < 0.01).
Fig. 5 is for passing through the brown eared pheasant chick of Euclidean system of distance cluster analysis 3 age in days FA content.Wherein, 1～No. 14 individual for survival, is for 15～No. 30 dead individual.Figure divides middle and upper part 19 individualities for high dead (HM) group, and all the other 11 individualities are low death (LM) group.
Fig. 6 is for passing through the brown eared pheasant chick of Euclidean system of distance cluster analysis 8 age in days FA content.Wherein, 1～No. 14 individual for survival, is for 15～No. 30 dead individual.It is HM group that figure divides middle and upper part 12 individualities, and all the other 18 individualities are LM group.
Fig. 7 is for passing through the brown eared pheasant chick of Euclidean system of distance cluster analysis 13 age in days FA content.Wherein, 1～No. 14 individual for survival, is for 15～No. 30 dead individual.It is LM group that figure divides middle and upper part 18 individualities, and all the other 12 individualities are HM group.
Embodiment
The experimental technique using in following embodiment if no special instructions, is conventional method.
Material, reagent etc. used in following embodiment, if no special instructions, all can obtain from commercial channels.
One, brown eared pheasant chick grouping to be measured and raising
Taking the brown eared pheasant chick of in May, 2011～September Pang Quan ditch nature reserve artificial incubation as research object.Between 24～June 12 May, hatch 93 of chick, incubation rate 90% from 103 pieces of ovum of 9 nests in field.30 of the chick of hatching birth on May 29 are chosen in the mensuration research of IgA content, from 3 nests, compile respectively as A group (14), B group (7), C group (9).When chick 2 age in days, carry out colored steel loop mark, the heavy 0.8g of steel loop, changes 1 time for after this every 30 days.
Two, the collection of excrement sample
On 31 days～September 1 May in 2011, adopt the method for non-damage technology, gathers the brown eared pheasant chick fresh excretas (feces collection weight > 0.5g) of 30 color ring marks with disposable glove, tweezers.In 2～35 age in days stages of chick, it is individual that sample collection uses indoor focus to observe, and gathers one to one fresh excreta.After chick 35 ages in days, breeding cage area alive is wide on a large scale, uses the remote focus of telescope to observe, after target individual defecation, and the fresh excrement sample of positioning acquisition.
Taking nest as unit gathers brown eared pheasant chick fresh excreta, selection of time 8:00～12:00 in the morning, after gathering, excrement sample packs receiving flask into, mark date, individual numbering, and put at once mobile icebox, after every day end-of-job in DEG C preservation of special refrigerator-freezer-20.Gather taking 5 days as one-period, after 60 ages in days because of weather reason, all prolongations of cycle.Research has gathered 18 cycles, collects altogether 338 parts, fresh excreta sample.
Three, the extraction of excrement sample
Chicken immunoglobulin extract: the phosphate buffer of the 0.01M that contains 0.05% polysorbas20 (volumn concentration), pH7.2; The solvent of the phosphate buffer of described 0.01M is water, and solute and concentration thereof are as follows: NaCl8.5g/L, Na
2hPO
42.2g/L and KH
2pO
40.3g/L.
Extract immunoglobulin in brown eared pheasant chick excrement sample: get the fresh excrement of 0.5g and be placed in 10ml centrifuge tube, add 5ml Chicken immunoglobulin extract, vortex instrument rotation extraction 10min, the centrifugal 20min of 2000r/min, get supernatant 1ml in 1.5ml centrifuge tube, the centrifugal 20min of 10000r/min again, gets supernatant in 1.5ml centrifuge tube,-20 DEG C of preservations, to be measured.
Four, the mensuration of excrement sample IgA
The content of immunoglobulin A (IgA) in enzyme linked immunosorbent assay quantitative assay above-mentioned steps three gained supernatant to be measured, thus calculate the content of IgA in Unit Weight ight soil.Use Chicken IgA ELISA Quantitation Set (E30-103, Bethyl Laboratories, and ELISA Starter Accessory Kit(E101 USA), Bethyl Laboratories, USA), provide handbook to prepare ELISA check-out console according to supplier, by microplate reader (Multiskan MK3, Thermo Scientific, USA) under 450nm light wave, measure the content of IgA in above-mentioned steps three gained supernatant to be measured.Concrete operations are as follows:
(1) coated antibody: to ELISA Plate (Maxisorp, Nunc, Denmark) in every hole, add 100 μ l IgA coated antibody (A30-103A, Bethyl Laboratories, USA) solution (dilutes according to volume ratio 1:100 with being coated with buffer solution (E107, Bethyl Laboratories, USA), pH9.6), hatch 1h.
(2) washing: use is washed every hole under plate machine (Wellwash 4 MK2, Thermo Scientific, USA) room temperature and added 400 μ l washing lotions (E106, Bethyl Laboratories, USA) to wash, and washes altogether 5 times, pats dry.
(3) sealing: every hole adds 200 μ l sealing buffer solutions (E104, Bethyl Laboratories, USA), hatches 30min.
(4) washing: same to step (2).
(5) standard items dilution and application of sample: add chicken IgA serum standard panel (RS10-102 in every hole, Bethyl Laboratories, USA) (concentration is followed successively by 1000.00ng/mL, 500.00ng/mL, 250.00ng/mL, 125.00ng/mL, 62.50ng/mL, 31.25ng/mL, 15.60ng/mL to gradient dilution liquid, with E106 washing lotion (the Bethyl Laboratories of polysorbas20 that adds volume fraction 0.05%, USA) dilute) or above-mentioned steps three gained supernatant to be measured, every hole 100 μ l, hatch 1h.
(6) washing: same to step (2).
(7) add enzyme labelled antibody: every hole adds the IgA enzyme labelled antibody (A30-103P of 100 μ l, Bethyl Laboratories, USA) dilution (is used E106 washing lotion (the Bethyl Laboratories of the polysorbas20 that adds volume fraction 0.05%, USA) dilute, dilution ratio is 1:100000), hatch 1h.
(8) washing: same to step (2).
(9) colour developing: every hole adds 100 μ l zymolytes (E102, Bethyl Laboratories, USA), and lucifuge is hatched 15min, after colour developing, solution colour is blue.
(10) stop: every hole adds the stop buffer (E115, Bethyl Laboratories, USA) of 100 μ l, cessation reaction, now blueness becomes yellow.
(11) measure: with blank hole (blank hole is without step (5) and step (7)) zeroing, use microplate reader (Multiskan MK3, Thermo Scientific, USA) the each hole of analyzing and testing absorbance (OD value) under 450nm light wave.Mensuration should be carried out in 15min adding after stop buffer.
Attention: above all operations carries out at 23 ± 1 DEG C.
The OD450 value (table 1) recording according to application of sample concentration value (ng/mL) and each hole of gauge orifice, production standard curve map (Fig. 1), according to calibration curve equation equation: y=2.815/[1+ (x/609.692)
-1.152]-0.015(R
2=0.9994), obtain the concentration (ng/mL) of IgA in sample well (above-mentioned steps three gained supernatant to be measured).According to formula: concentration (ng/ml) × 10ml/g of IgA in the content of IgA (ng/g)=above-mentioned steps three gained supernatant to be measured in every gram of fresh excreta, converts and obtains the amount (ng/g) of IgA in every gram of fresh excreta.
The application of sample concentration value (ng/ml) of table 1 gauge orifice and the OD value that each hole records
Elisa plate quantitatively detects, measurement range: 15.6ng/ml～1000ng/ml; Sensitivity: 7.3ng/ml; The parameter that specificity: IgA and IgG, IgM, the equal < 0.01%(of IgE cross reaction kit supplier provide).In addition, the present inventor is to 338 parts of fecal specimens replications 3 times, and through calculating, the coefficient of variation in plate and between plate is respectively 4.2% and 4.8%.The above-mentioned enzyme linked immunosorbent assay of visible employing is carried out quantitative assay to fecal specimens, its credible result, and accuracy rate is high.
Five, statistical analysis
The data that experiment obtains are carried out data analysis by SPSS 20.0, Excel 2010, use sigmaplot 12.0 data to draw.Data result represents with the form of " Mean scholar SE ".The data of analyzing are all carried out normal distribution-test by single sample K-S inspection (One-Sample Kolmogorov-Smirnov Test).In group, data are used one-way analysis of variance (one-way ANOVA), relatively adopt independent sample T inspection (Independent-samples T test) between group, use these two kinds of inspections all to carry out homogeneity test of variance, and significance level arranges α=0.05.Chick ight soil immunoglobulin content adopts square Euclidean distance (Euclidean distance) to carry out hierarchial-cluster analysis (Systematic cluster analysis).
Six, experimental result
1, the content of brown eared pheasant chick ight soil IgA
Brown eared pheasant chick 3～95 age in days ight soil IgA(FA) quantitative assay result, maximum 9002.33ng/g, minimum of a value 309.74ng/g, the FA mean value of each age in days is shown in Fig. 2.There are 4 crests in FA mean value curve, is respectively 13 ages in days (1609.24 ± 114.14ng/g), 28 ages in days (5882.49 ± 451.54ng/g), 42 ages in days (2970.14 ± 456.50ng/g), 50 ages in days (1560.09 ± 595.66ng/g).When chick 20 age in days, occur first dead individuality, before it is dead, the content of FA is starkly lower than colony's mean value.Dead 10 of 20～30 ages in days, dead 2 of 30～40 ages in days, dead 4 of 40～50 ages in days.
2, ight soil IgA between chick survival and dead individuality
When 30 chick 95 ages in days, 16 of Died Of Diseases, lethality is 53.3%.Dead individual ight soil IgA(DFA) content and the individual ight soil IgA(SFA of surviving) the vertical scatter diagram of content, see Fig. 3.The average content of 3 age in days DFA is 727.0 ± 53.3ng/g, and the average content of 3 age in days SFA is 1115.6 ± 87.8ng/g.Before chick 50 ages in days, the average content of SFA and DFA is all higher than its 3 age in days level, and after 50 ages in days, the average content of SFA is lower than 3 age in days levels.DFA and the SFA of 3～50 ages in days (test point is respectively 3 ages in days, 8 ages in days, 13 ages in days, 18 ages in days, 23 ages in days, 28 ages in days, 33 ages in days, 38 ages in days, 42 ages in days, 48 ages in days, 50 ages in days) carry out independent sample t inspection, result is as Fig. 4, show that both all exist utmost point significant difference (df=28 at 3 ages in days, 8 ages in days, 13 ages in days, 18 ages in days, P < 0.01), there is significant difference in 33 ages in days (df=18, P < 0.05).All the other ages in days are all without significant difference.
3, chick ight soil IgA and lethality
By Euclidean system of distance cluster analysis chick 3 ages in days, 8 ages in days, 13 age in days FA content, result can be divided into high mortality group (HM) and low actual group (LM).Chick 3 age in days FA content, are shown in Fig. 5, and the FA content of HM group is at 436.47-833.67ng/g, and lethality is 78.94%(15/19), the FA content of LM group is at 1132.61-1642.40ng/g, lethality is 9.10%(1/11).Chick 8 age in days FA content, are shown in Fig. 6, and the FA content of HM group is at 647.41-1195.61ng/g, and lethality is 83.33%(10/12), the FA content of LM group is at 1363.92-2337.40ng/g, lethality is 33.33%(6/18).Chick 13 age in days FA content, are shown in Fig. 7, and the FA content of HM group is at 558.38-1298.07ng/g, and lethality is 75.00%(9/12), the FA content of LM group is at 1556.32-2611.95ng/g, lethality is 33.33%(5/18).
The raising of embodiment 2, brown eared pheasant the test of hiving off of brooding
The brown eared pheasant of 2011 and 2012 2 years is raised the test of hiving off of brooding
One, the raising of 2～4 Japanese instar chicklings
Experimental animal and grouping: 60 come from the brown eared pheasant chick of 2～4 age in days of Pang Quan ditch nature reserve artificial incubation, are divided at random two large groups, 15 of 45 of experimental group and control groups.
Experimental group: according to the method described in embodiment 1, the content of the brown eared pheasant chick of determination experiment group ight soil IgA.According to assay result, with reference to passing through the result of the brown eared pheasant chick of Euclidean system of distance cluster analysis 3 age in days ight soil IgA content in embodiment 1 step 63,60 brown eared pheasant chick of 2～4 age in days are divided into following three groups, isolated rearing between group:
Group A: in ight soil, the content of immunoglobulin A is lower than waiting to educate brown eared pheasant chick, totally 20 described in 834ng/g;
Group B: in ight soil, the content of immunoglobulin A is higher than waiting to educate brown eared pheasant chick, totally 16 described in 1133ng/g;
Group C: in ight soil the content of immunoglobulin A between 834ng/g and 1133ng/g described in wait to educate brown eared pheasant chick, totally 9.
Control group: do not carry out the mensuration of ight soil IgA content, at random by the isolated rearing of 15 brown eared pheasant chick of 2～4 age in days:
Chick to experimental group and control group is raised, and when chick 3～35 age in days, the hen band of being raised by protection zone is young, taking group as unit, raises respectively in breeding chickling chamber, and sufficient chicken feed and identical room temperature are provided.Every day, 10:00-15:00, carried out isolated rearing between group by each group chick in outdoor breeding cage.After chick 35 ages in days, each group all puts into breeding cage and carries out isolated rearing between group, raises chick always and becomes after sub-one-tenth chicken (i.e. 90 ages in days), the lethality of brown eared pheasant in statistical experiment group and control group.In whole feeding process, need to keep the consistent of experimental group and control group condition.
Result is as shown in table 2, under identical raising condition, and final dead 12 of 15 brown eared pheasants of control group, its lethality is up to 80.0%; And 45 brown eared pheasants of experimental group, A organizes dead 17, each dead 2 of B group and C group, and statistics somatic death rate is only 46.7%, far below control group, and both significant differences, there is statistical significance.
The table 2 difference method of brooding is raised the statistics of the brown eared pheasant chick of 3 age in days lethality
Two, the raising of 7～9 Japanese instar chicklings
Experimental animal and grouping: 60 come from the brown eared pheasant chick of 7～9 age in days of Pang Quan ditch nature reserve artificial incubation, are divided at random two large groups, 15 of 45 of experimental group and control groups.
Experimental group: according to the method described in embodiment 1, the content of the brown eared pheasant chick of determination experiment group ight soil IgA.According to assay result, with reference to passing through the result of the brown eared pheasant chick of Euclidean system of distance cluster analysis 8 age in days ight soil IgA content in embodiment 1 step 63,45 brown eared pheasant chick of 7～9 age in days are divided into following three groups, isolated rearing between group:
Group A: in ight soil, the content of immunoglobulin A is lower than waiting to educate brown eared pheasant chick, totally 13 described in 1196ng/g;
Group B: in ight soil, the content of immunoglobulin A is higher than waiting to educate brown eared pheasant chick, totally 19 described in 1363ng/g;
Group C: in ight soil the content of immunoglobulin A between 1196ng/g and 1363ng/g described in wait to educate brown eared pheasant chick, totally 13.
Control group: do not carry out the mensuration of ight soil IgA content, at random by the isolated rearing of 15 brown eared pheasant chick of 7～9 age in days:
Chick to experimental group and control group is raised, and when chick 7～35 age in days, the hen band of being raised by protection zone is young, taking group as unit, raises respectively in breeding chickling chamber, and sufficient chicken feed and identical room temperature are provided.Every day, 10:00-15:00, carried out isolated rearing between group by each group chick in outdoor breeding cage.After chick 35 ages in days, each group all puts into breeding cage and carries out isolated rearing between group, raises chick always and becomes after sub-one-tenth chicken (i.e. 90 ages in days), the lethality of brown eared pheasant in statistical experiment group and control group.In whole feeding process, need to keep the consistent of experimental group and control group condition.
Result is as shown in table 3, under identical raising condition, and final dead 12 of 15 brown eared pheasants of control group, its lethality is up to 80.0%; And 45 brown eared pheasants of experimental group, A organizes dead 10, and B organizes dead 3, and C organizes dead 4, and statistics somatic death rate is only 37.8%, far below control group, and both significant differences, there is statistical significance.
The table 3 difference method of brooding is raised the statistics of the brown eared pheasant chick of 8 age in days lethality
Three, the raising of 12～14 Japanese instar chicklings
Experimental animal and grouping: 60 come from the brown eared pheasant chick of 12～14 age in days of Pang Quan ditch nature reserve artificial incubation, are divided at random two large groups, 20 of 45 of experimental group and control groups.
Experimental group: according to the method described in embodiment 1, the content of the brown eared pheasant chick of determination experiment group ight soil IgA.According to assay result, with reference to passing through the result of the brown eared pheasant chick of Euclidean system of distance cluster analysis 13 age in days ight soil IgA content in embodiment 1 step 63,45 brown eared pheasant chick of 12～14 age in days are divided into following three groups, isolated rearing between group:
Group A: in ight soil, the content of immunoglobulin A is lower than waiting to educate brown eared pheasant chick, totally 15 described in 1298ng/g;
Group B: in ight soil, the content of immunoglobulin A is higher than waiting to educate brown eared pheasant chick, totally 19 described in 1556ng/g;
Group C: in ight soil the content of immunoglobulin A between 1298ng/g and 1556ng/g described in wait to educate brown eared pheasant chick, totally 11.
Control group: do not carry out the mensuration of ight soil IgA content, at random by the isolated rearing of 15 brown eared pheasant chick of 12～14 age in days:
Chick to experimental group and control group is raised, and when chick 12～35 age in days, the hen band of being raised by protection zone is young, taking group as unit, raises respectively in breeding chickling chamber, and sufficient chicken feed and identical room temperature are provided.Every day, 10:00-15:00, carried out isolated rearing between group by each group chick in outdoor breeding cage.After chick 35 ages in days, each group all puts into breeding cage and carries out isolated rearing between group, raises chick always and becomes after sub-one-tenth chicken (i.e. 90 ages in days), the lethality of brown eared pheasant in statistical experiment group and control group.In whole feeding process, need to keep the consistent of experimental group and control group condition.Experiment repeats 3 times, results averaged.
Result is as shown in table 4, under identical raising condition, and final dead 11 of 15 brown eared pheasants of control group, its lethality is up to 73.3%; And 45 brown eared pheasants of experimental group, A organizes dead 11, and B organizes dead 2, and C organizes dead 3, and statistics somatic death rate is only 35.6%, far below control group, and both significant differences, there is statistical significance.
The table 4 difference method of brooding is raised the statistics of the brown eared pheasant chick of 12～14 age in days lethality
The result of cumulated volume embodiment step 1 to three; visible employing is provided by the present invention will wait to educate brown eared pheasant chick; according to the isolated rearing of dividing into groups of the content of immunoglobulin A in ight soil; compared with the control group of random packet; for whole chick colony; this feeding manner greatly reduces the lethality of chick, and this protection artificial breeding for the brown eared pheasant of endangered species is significant.
Claims (1)
1. the brown eared pheasant method of brooding, for by 2～14 ages in days wait educate brown eared pheasant chick according to the isolated rearing of dividing into groups of the content difference of immunoglobulin A in ight soil, it is characterized in that: described method is any in following (1)～(3):
(1) 2～4 ages in days are waited to educate brown eared pheasant chick and are divided into following three groups, isolated rearing between group:
Group A: in ight soil, the content of immunoglobulin A is lower than waiting to educate brown eared pheasant chick described in 834ng/g;
Group B: in ight soil, the content of immunoglobulin A is higher than waiting to educate brown eared pheasant chick described in 1133ng/g;
Group C: in ight soil the content of immunoglobulin A between 834ng/g and 1133ng/g described in wait to educate brown eared pheasant chick;
(2) 7～9 ages in days are waited to educate brown eared pheasant chick and are divided into following three groups, isolated rearing between group:
Group A: in ight soil, the content of immunoglobulin A is lower than waiting to educate brown eared pheasant chick described in 1196ng/g;
Group B: in ight soil, the content of immunoglobulin A is higher than waiting to educate brown eared pheasant chick described in 1363ng/g;
Group C: in ight soil the content of immunoglobulin A between 1196ng/g and 1363ng/g described in wait to educate brown eared pheasant chick;
(3) 12～14 ages in days are waited to educate brown eared pheasant chick and are divided into following three groups, isolated rearing between group:
Group A: in ight soil, the content of immunoglobulin A is lower than waiting to educate brown eared pheasant chick described in 1298ng/g;
Group B: in ight soil, the content of immunoglobulin A is higher than waiting to educate brown eared pheasant chick described in 1556ng/g;
Group C: in ight soil the content of immunoglobulin A between 1298ng/g and 1556ng/g described in wait to educate brown eared pheasant chick.
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| KR100624012B1 (en) * | 2001-04-04 | 2006-09-15 | 주식회사 녹십자홀딩스 | Kit for Diagnosing Non-pregnancy, and Method for Diagnosing Non-pregnancy Using the Same |
| CN102037904A (en) * | 2009-10-22 | 2011-05-04 | 姜哲 | Poultry brooding method using fermentation bed with tenebrio molitor frass thereon and dedicated device thereof |
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