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1.
In a previous study, we reported a new method for counting quail blood cells. After quail blood cells were stained with fluorescent lipophilic dye (DiOC6(3)), absolute counts of erythrocytes, granulocytes, and monocytes were obtained by means of flow cytometry (FC). The FC method has the potential for application to avian blood cells count; however, the method was unable to distinguish between lymphocytes and thrombocytes. In the present study, we improved the FC method to obtain separate counts of lymphocytes using DiOC5(3). After quail blood cells were stained with DiOC5(3), the cells were measured with FC. Each blood cell type was distinguished by means of their typical FL-1 (green fluorescence) and SSC (side scatter). Absolute numbers of erythrocytes, granulocytes, monocytes and lymphocytes in whole blood were obtained. The improved FC analysis worked equally well with chicken (Gallus gallus) and goose (Anser cygnoides) blood.  相似文献   

2.
OBJECTIVE: To evaluate the potential usefulness of 2 flow cytometric methods for determination of differential cell counts in feline bone marrow. SAMPLE POPULATION: 10 bone marrow specimens from client-owned cats. PROCEDURE: Bone marrow specimens were stained with 3,3'-dihexyloxacarbocyanine iodide (DiOC6) and evaluated by use of flow cytometry. Differential counts were also determined by analysis of scatterplots of forward-angle versus side-angle light scatter of unstained specimens, obtained by use of flow cytometry (scatterplot method). Results of both flow cytometric methods were compared with differential cell counts determined by manually counting 1,000 cells on slides of Wright-stained smears. RESULTS: Staining with DiOC6 resulted in identification of mature and immature erythroid and myeloid cells and lymphocytes. Use of the scatterplot method resulted in identification of mature and immature erythroid and myeloid cells and metamyelocytes. However, to identify lymphocytes by use of the scatterplot method, bone marrow specimens were first labeled with an anti-major histocompatability class-II antibody. Comparison of results of the scatterplot method with manual counts yielded higher correlation coefficients and more similar mean values than did comparison of results of the DiOC6 method. Conclusions and Clinical Relevance: The scatterplot method provided more accurate and precise results than the DiOC6 method for determination of bone marrow differential cell counts in cats by use of flow cytometry. When combined with fluorescent labeling of lymphocytes, the scatterplot method has potential to provide rapid semiquantitative assessment of bone marrow differential cell counts in cats.  相似文献   

3.
Abstract: Three flow cytometric techniques were evaluated for determination of differential cell counts on canine clinical bone marrow specimens. Techniques included staining bone marrow specimens with 2'7'-dichlo-rofluorescein (DCF) or 3,3'-dihexyloxacarbocyanine iodide (DiOC6) and evaluation of forward-angle light scatter vs. side-angle light scatter plots. Flow cytometric evaluation of bone marrow cells stained with DCF failed to separate bone marrow cells into distinct cell populations. Staining with DiOC6 resulted in separation of bone marrow cells into populations of mature and immature erythroid cells, mature and immature myeloid cells, and lymphocytes. The scatter plot method resulted in identification of mature and immature erythroid cells, immature myeloid cells, metamyelocytes, and bands and segmenters. Lymphocytes could not be differentiated from mature erythroid cells by the scatter plot method. When the results of the DiOC6 method and the scatter plot method were compared with manual bone marrow differential cell counts, the scatter plot method had more similar mean values and higher correlation coefficients. The scatter plot method has the potential of providing rapid semiquantitative assessment of bone marrow differential cell counts in dogs for specimens that contain low numbers of lymphocytes.  相似文献   

4.
The effect of selection for high stress response on adrenocortical function was examined by measuring the corticosterone response of adrenocortical cells isolated from random-bred Japanese quail and quail selected for high serum corticosterone response to immobilization (high-stress). Highly enriched adrenocortical cells were incubated with various concentrations of ACTH1-24 (ACTH), 8-bromo-cyclic AMP (8Br-cAMP) and pregnenolone for 2 hr. Corticosterone production was measured by radioimmunoassay. Basal corticosterone production values by cells from random-bred and high-stress birds were not different. In contrast, the average maximal ACTH- and 8Br-cAMP-induced corticosterone production by cells from high-stress quail was 89% greater than that of cells from random-bred quail. However, the average pregnenolone-supported corticosterone production by cells from high-stress birds was 34% less than that of cells from random-bred birds. Thus, the data suggest that although random-bred quail cells had a greater potential capacity for corticosterone production, high-stress quail cells had a greater ability to couple ACTH, ACTH-transmembrane-signaling factors and subsequent second messengers with the available steroidogenic enzyme pool. The magnitude of the differences in function between cells from high-stress and random-bred birds was greater for female cells compared to male cells. In addition to differences in cellular function, there were also differences in adrenal and relative adrenal weights between random-bred and high-stress quail. The average, adrenal and relative (mg/100 g body weight) adrenal weights of high-stress quail were 14-16% greater than those of random-bred quail. It is concluded that the enhanced serum corticosterone response of the high-stress quail line is, in part, due to an increase in relative adrenal weight and an increase in adrenocortical cell responsiveness to ACTH.  相似文献   

5.
Production of quail-chick chimaeras by blastoderm cell transfer   总被引:2,自引:0,他引:2  
1. Quail-chick chimaeras were produced by injecting dissociated quail blastoderm cells into chick embryos. 2. Quail blastoderms were removed from the yolk and the cells were dispersed by trypsin treatment or pipetting. The cell suspension (1 to 5 microliters) was injected into the subgerminal cavity of unincubated chick embryos. The chick embryos were then cultured in recipient eggshells. 3. Quail blastoderm cells injected into the chick embryos adhered to the chick embryonic cells. The rates of hatching were 8.6% (38 chicks from 441 eggs) and 40.3% (48 chicks from 119 eggs) when the volumes of the cell suspension injected were 3 to 5 microliters and 1 microliter, respectively. 4. Seven out of 86 hatched birds were clearly identified as being chimaeric because part of the feather colouring was of quail specificity. In addition to these chimaeric birds, there were 8 chimaeric embryos which died before hatching. The distribution patterns of the quail feathers were varied among the chimaeric birds and embryos. 5. This technique provides a basis for the investigation of chick embryo cryopreservation, genetic transformation and analysis of cell lineage of chickens.  相似文献   

6.
The aim of the present study was to examine whether chicken semen can be substituted for quail semen to conduct in vitro experiments on fertilization. Chicken spermatozoa was incubated with the inner perivitelline layer (IPL) isolated from the largest follicle in the quail ovary under in vitro conditions. The perforation of chicken and quail spermatozoa were assessed by counting the number of all visible holes in the pieces of IPL. No difference was found in the number of holes formed by the chicken sperm and quail IPL interaction compared with that between intraspecies. In addition, the number of holes in the IPL was significantly increased with the increase in sperm concentration from 1 × 106 to 8 × 106 sperm/mL (P < 0.05). Interestingly, after treatment of chicken spermatozoa with 2.5 mg/mL of the solubilized quail IPL followed by incubation with the intact chicken IPL, the number of holes in the intact chicken IPL was significantly decreased as compared with that of spermatozoa treated without the solubilized IPL (P < 0.05). This indicates that sperm receptors in the solubilized quail IPL and binding ligands on the chicken spermatozoa would find each other and bind, forming complexes and these complexes blocked the interaction between chicken spermatozoa and intact chicken IPL. These results show that: (i) chicken spermatozoa possess the penetrability into quail IPL; and (ii) a high degree of affinity via the receptor interactions exists between chicken spermatozoa and quail IPL. Therefore, it appears that the substitution of chicken semen for quail semen is possible to use as an in vitro technique to examine the sperm‐IPL interaction during fertilization in quail.  相似文献   

7.
The development of preimplantation mammalian embryos in vitro is less than optimal. Follicular fluid and cumulus cells have both been used independently, to improve preimplantation embryo quality in culture. This study was undertaken to evaluate the influence of a cumulus cell monolayer in human follicular fluid on mouse early embryo development in vitro. One-cell embryos were obtained from NMRI mice after superovulation with eCG and hCG. Cumulus cells were prepared from mouse egg-cumulus mass. These cells were separated from red blood cells using a Percoll gradient. Follicular fluid was collected from patients undergoing an IVF program during oocyte pick-up. The cumulus cell monolayer was prepared in follicular fluid (FC) and Ham's F10 (HC). Mouse one-cell embryos were cultured in FC, HC, Ham's F10 (HF) and follicular fluid (FF) for 120 h. Only 10.5% of embryos passed the two-cell block in HF. However, the proportions of embryos passing the two-cell block were 23.1%, 21.4% and 68.5% in FF, HC, and FC treatments, respectively; which were significantly different from HF (p<0.05). The differences between FC and the two other treatments were also significant (p<0.001). In FC, 33.7% of one-cell embryos continued to grow to the blastocyst stage whereas only 2.1% and 1.9% of one-cell embryos in FF and HC reached this stage and no embryos developed to blastocyst in HF. The proportion of blastocysts in FC was significantly higher than all other treatments (p<0.001). It can be concluded that follicular fluid and cumulus cells in monolayer form synergistically improve the early embryo culture condition.  相似文献   

8.
This study was conducted to investigate the effect of sex, age, number of bronchoalveolar lavages and method of quantitation on the number of bronchoalveolar cells in rats. Forty Long Evans rats were divided into four age-sex subgroups of ten animals each. Nine consecutive bronchoalveolar lavages were done in every rat and the number of bronchoalveolar cells/mL in lavages 1-3 (L1), 4-6 (L2) and 7-9 (L3) were determined by hemocytometer and electronic cell counts (Coulter Counter). The sex or age of the rats did not show a significant effect (p less than 0.05) in the number of bronchoalveolar cells recovered from the lungs; however, there was a significant difference (p less than 0.05) in the number of cells/mL among lavages 1-3, 4-6 and 7-9 (L1 approximately equal to L2 greater than L3). A discrepancy of approximately 8% in the counts of bronchoalveolar cells was found between the hemocytometer and the Coulter Counter; however, these two methods of cell quantitation showed a significant (p less than 0.01) positive correlation (r = 0.89). No significant (p greater than 0.05) differences were found in the percentage of fluid recovery (overall mean = 94.5%) among lavages L1, L2 and L3. It was concluded that the electronic cell counting of bronchoalveolar cells is as reliable as manual counting. Although sex or age did not significantly affect the number of cells recovered from the lung, caution should be used in the number of lavages done per rat since this variable may significantly affect the results.  相似文献   

9.
The effects of reticuloendotheliosis virus (REV) infection were studied using an experimental model in Japanese quail during 2 consecutive generations. The REV used in this study (APC-566) was isolated from Attwater's prairie chickens (Tympanuchus cupido attwateri). Tumors were induced by APC-566 as early as 6 wk of age. Mortality was significantly higher in groups of quail with a higher frequency of REV infection. Egg production, hatchability, and fertility rates decreased in infected quail as compared with uninfected control quail. The BW of infected quail were significantly reduced at 8 wk of age in the first generation of infected quail (breeders) and at 3 and 6 wk of age in the second generation (quail broilers) compared with uninfected quail.  相似文献   

10.
The biologically active form of vitamine D(3) [1alpha,25(OH)(2)D(3)] has recently been described not only to influence bone metabolism but also to exert immunomodulating activities, which may have an impact on bone formation/resorption as well. In this study, we analysed the effects of 1alpha,25(OH)(2)D(3) on the cytokine pattern of porcine bone marrow-derived cells from piglets aged 1-3 weeks. After culture for 1 week, the number of osteoclasts was determined, with tartrate-resistant acid phosphatase (TRAP)-positive, multinucleated cells being considered osteoclasts. Cultured bone marrow cell-derived mRNA was subjected to semiquantitative RT-PCR specific for a panel of porcine cytokines (IL-1alpha, IL-6, IL-8, IL-10, and TNF-alpha). In addition, an immunofluorescence analysis using anti-porcine mAbs specific for IL-1beta, IL-2, IL-4, IL-6, IL-12, TNF-alpha, and IFN-gamma was performed. In order to prove the existence of a porcine homologue of the receptor activator of NF-kappaB ligand (RANKL) bone marrow cell- as well as porcine white blood cell-derived mRNA was investigated by RT-PCR using primer pairs specific for murine RANKL. Cell culture supernatant was analysed for soluble RANKL by means of an ELISA designed for quantification of human RANKL. By means of RT-PCR, expression of IL-1alpha, IL-6, IL-8, IL-10 and TNF-alpha mRNA could be found in cells cultured with and without 1alpha,25(OH)(2)D(3). Immunofluorescence analysis revealed that IL-1, IL-6, and TNF-alpha were produced by both stromal cells and osteoclasts. Besides its known osteoclastogenic effects, 1alpha,25(OH)(2)D(3) tended to downregulate the respective cytokines, but significantly upregulated RANKL expression. The homology between the porcine RANKL-specific sequence and the corresponding human RANKL sequence was 79%. The data found support the idea that porcine bone marrow cell cultures may provide a suitable alternative to murine systems in human osteological research.  相似文献   

11.
Manganese deficiency results in neurological and skeletal defects, together with ultrastructural disarrangement of the retina in rats. Wild birds show a range of Mn concentrations in their tissues, including the liver, raising the possibility of Mn-related disorders in the wild. Electroretinography (ERG) provides a useful noninvasive approach to evaluate visual function. This method is especially useful in birds, as objective analysis of them is very difficult, while they have well-developed vision. In this study, we carried out a convenient and reliable ERG recording using a contact lens electrode with a built-in light source (LED electrode) of Japanese quail (Coturnix coturnix japonica) fed a Mn-deficient diet. After 10 min light adaptation, single-flash and flicker cone responses were reproducibly recorded to cause an intensity-dependent increase in amplitude of both a-wave and b-wave in single-flash ERG. Mn-deficient feeding markedly decreased the Mn concentration in the liver by almost half in 3 to 6 weeks, followed by body weight loss in 13 to 15 weeks. Implicit time of a-wave and b-wave cone response by single-flash stimulation was significantly delayed in quail with a Mn depletion from 3 to 6 weeks. Every cone response of the Mn-deprived quail had a tendency to decrease amplitude. The ultrastructure of cone photoreceptor cells was disorganized by Mn deficiency, including changes in outer segment discs of photoreceptor cells. These results suggest the essential role of Mn in the integrity of the retinal function of birds.  相似文献   

12.
The purpose of this study was to accelerate the application of molecular markers in genetic breeding and other aspects of Black quail to develop EST-SSR markers for Black quail.100 Black quail were randomly chosen and blood samples were collected to develop EST-SSR new markers and determined the genetic polymorphism using bioinformatics.The results showed that there were 6 EST-SSR markers in the Black quail, the number of observed alleles in EST-SSR was 2 to 6.The average PIC of 6 EST-SSR markers in the Back quail was 0.4962 and the average He was 0.5759.Markers P1 and P2 were highly polymorphic markers among 6 EST-SSR and the rest EST-SSR markers were moderately polymorphic markers.These developed markers could be used for genetic diversity analysis of Black quail and the EST-SSR markers had some relationship with the carbohydrate metabolism, synthesis of melanin in Black quail, but the mechanism needed the further analysis.  相似文献   

13.
1. The effect of increasing dietary selenium (Se) on production performance and immune responses in growing (0 to 6 weeks) Japanese quail was investigated. 2. One-day-old chicks (240) were randomly selected and divided into 12 groups with 20 chicks in each group (3 dietary treatments x 4 replicates). The basal diet contained 0.2 mg Se/kg and the two experimental diets were supplemented with 0.5 and 1.0 mg Se/kg. 3. Body weight gain, food intake and food conversion ratio and mortality were not affected by Se supplementation. 4. On d 28, antibody responses to inoculated sheep red blood cells were determined. Antibody titres were significantly higher after feeding the two Se-supplemented diets. 5. During week 4, the response to intradermally injected phytohaemagglutinin, an index of the in vivo cell-mediated immune response, was shown to be increased in the groups fed on the Se-supplemented diets. 6. After 6 weeks, the relative weights of the bursa of Fabricius and thymus were greater in the chicks given the Se-supplemented diets but there was no effect on the relative weight of spleen and liver. 7. It is concluded that supplementing the diet with Se has a beneficial effect on immune responses but does not affect production performance in growing Japanese quail.  相似文献   

14.
本研究旨在加速分子标记在黑羽鹌鹑遗传育种等方面的应用,开发黑羽鹌鹑的EST-SSR标记。随机选取100只黑羽鹌鹑,心脏采血2mL,采用生物信息学方法开发黑羽鹌鹑的EST-SSR新标记,并检测EST-SSR标记在黑羽鹌鹑群体中的多态性。结果显示,在黑羽鹌鹑中成功的开发出了6个EST-SSR标记,EST-SSR标记检测到的观测等位基因数为2~6个,6个EST-SSR标记在黑羽鹌鹑的平均多态信息含量(PIC)为0.4962,平均期望杂合度(He)为0.5759。6个EST-SSR标记中P1和P2个为高度多态标记,其他4个EST-SSR为中度多态标记。新开发的6个EST-SSR标记可用于黑羽鹌鹑的遗传多样性分析,6个EST-SSR标记与鹌鹑的糖类代谢、黑色素的合成等有关系,但具体的机理还有待进一步分析。  相似文献   

15.
M Ferris  W J Bacha 《Avian diseases》1984,28(1):179-182
A method for the direct counting of chicken heterophils and eosinophils using Phloxine B stain and a hemacytometer is described. A solution of 0.1% Phloxine B in 50% propylene glycol stained heterophils and eosinophils obtained from peripheral blood. The cytoplasm of heterophils became completely stained, whereas that of the eosinophils stained incompletely, thereby enabling these cells to be distinguished easily from each other. Identification of cell type was confirmed by comparing cell counts prepared from blood smears stained with Wright's stain with wet mounts (hemacytometer preparations) stained with Phloxine B.  相似文献   

16.
We investigated vascular access ports for feline blood donation. Eight cats were anesthetized for conventional blood collection by jugular venipuncture at the beginning and end of the study. In-between conventional collections, vascular access ports were used for collection with or without sedation every 6 to 8 wk for 6 mo. Ports remained functional except for one catheter breakage, but intermittent occlusions occurred. Systolic blood pressure was lower during conventional collection. Behavioral abnormalities occurred during 3 port collections. Packed red cells prepared from collected blood were stored at 4°C for 25 d and assessed for quality pre- and post-storage. With both collection methods, pH and glucose level declined, and potassium level, lactate dehydrogenase activity and osmotic fragility increased. There were no differences between methods in pre-storage albumin and HCO(3)(-) levels, and pre and post-storage hematocrit, lactate dehydrogenase activity, and glucose and potassium levels. Pre-storage pH and pCO(2) were higher with conventional collection, and pre- and post-storage osmotic fragility were greater with port collection. One port became infected, but all cultures of packed red cells were negative. Tissue inflammation was evident at port removal. In a second study of conventional collection in 6 cats, use of acepromazine in premedication did not exacerbate hypotension. The use of vascular access ports for feline blood donation is feasible, is associated with less hypotension, and may simplify donation, but red cell quality may decrease, and effects on donors must be considered.  相似文献   

17.
Day-old quails experimentally infected with Marek's disease (MD) virus of quail origin developed lymphoid tumors. The severity of the disease increased considerably with serial passage. Tumor transplants could be made with cells derived from gross tumors in skeletal muscles, spleen cells, and blood from MD-affected quails. After five to six serial transplants, the tumor could not be transplanted further. Marek's disease tumor-associated surface antigen (MATSA) was demonstrated in lymphoid cells of spleen and peripheral blood lymphocytes of MD-affected quails. The MATSA of quail differed from the MATSA of chicken. Chickens were susceptible to MD virus isolated and propagated in quails.  相似文献   

18.
鸡马立克氏病活疫苗免疫效力比较试验   总被引:1,自引:0,他引:1  
用HVT冻干苗、HVT细胞结合苗、CVI988细胞结合苗、SB1+FC126双价活疫苗、301B/1+FC126双价活疫苗和Z4+FC126双价活疫苗等6种鸡马立克氏病(MD)疫苗免疫SPF白来航鸡或普通伊莎鸡,用鸡马立克氏病病毒(MDV)强毒GA株、京-1血毒以及鸡马立克氏病超强毒vvMDV-Md5毒株分别攻击进行免疫效力比较试验。试验表明,MD单价苗的免疫效力强弱顺序依次是CVI988、HVT细胞结合苗和HVT冻干苗,这3种MD单价苗均能给免疫鸡群提供有效的免疫保护力。SB1+FC126、Z4+FC126和301B/1+FC126等3种MD双价苗免疫效力显著高于MD单价苗,均能给免疫鸡群提供较强的免疫保护力,并能有效地抵抗vvMDV-Md5毒株的致瘤作用。Z4+FC126和301B/1+FC126MD双价苗免疫效力无显著差异  相似文献   

19.
1. The aim of the present study was to investigate the effects on the eggs and hatchlings (up to 2 weeks post-hatch) of feeding a relatively large amount of so-called organic selenium to breeder quail. 2. Two groups of quail (3 families in each group consisting of 4 females and 1 male) were formed at the beginning of their reproductive period. The quail were fed on a commercial maize-based diet containing 0.096 mg/kg feed-derived selenium (Se), supplemented with 0.2 mg/kg selenite (control group) or 0.5 mg/kg organic selenium in the form of Sel-Plex (Alltech Ltd, USA) for 6 months. Eggs were collected at 6 months of age and Se in the egg yolk, egg white and shell was analysed. Five quail at 1, 7 and 14 d post-hatch were killed to provide samples of liver, brain, breast and leg muscles for Se analysis. After egg collection for analysis and incubation, adult quail were killed and liver, kidney, lung, brain, breast and leg muscles were collected for Se analyses. 3. Inclusion of high doses (0.5 mg/kg) of organic Se in the quail diet was associated with a significant increase in Se concentration in all tissues studied of adult quail as well as in egg yolk, egg albumin and eggshell. 4. Increased Se concentration in the quail egg was associated with increased Se concentration in the liver, breast and leg muscles and brain of newly hatched quail. This difference was shown to be significant for 2 weeks post-hatch. Therefore, it has been suggested that the maternal effect of dietary selenium can be seen beyond the hatching time and more emphasis should be given to this effect in future. 5. It was shown that it is possible to produce Se-enriched quail meat and eggs by adding organic selenium to the diet.  相似文献   

20.
(1) The nitrogen-corrected apparent metabolisable energy (AME(N)) content of solvent-extracted rapeseed and sunflower seed (un-decorticated) meals in relation to species (chicken, guinea fowl and quail) and dietary addition of feed enzymes (0 or 0.5 g/kg diet) was evaluated by a diet replacement method in a 3 x 2 factorial design. (2) The metabolism trial was conducted at two substitution levels (200 and 400 g/kg diet) of each meal with or without supplementation of commercial enzyme preparation in 6 individuals or 6 groups of cockerels, guinea fowls and quails. (3) The experimental diets were fed for a period of 12 d followed by a 3-d collection period during which total feed consumed and droppings output were quantitatively recorded. (4) The AME(N) values of rapeseed meal for cockerels, guinea fowls and quails were 8.4, 8.7 and 8.8 MJ/kg, respectively, while the corresponding values for sunflower seed meal were 6.1, 6.1 and 6.2 MJ/kg dry matter, without enzyme supplementation. (5) The AME(N) value of rapeseed meal did not improve with enzyme supplementation. However, AME(N) values of sunflower seed meal significantly increased with enzyme supplementation, from 6.1 to 6.5 MJ/kg dry matter. (6) Since AME(N) values of rapeseed meal and sunflower seed meal were similar in chicken, guinea fowl and quail, values reported for chicken could, therefore, be used for guinea fowl and Japanese quail.  相似文献   

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