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Development of an enzyme-linked immunosorbent assay to detect serum antibody to chicken anemia agent 总被引：3，自引：0，他引：3
D Todd D P Mackie K A Mawhinney T J Connor F McNeilly M S McNulty 《Avian diseases》1990,34(2):359-363
An enzyme-linked immunosorbent assay (ELISA) for the detection of antibodies to chicken anemia agent (CAA) has been developed. This test utilizes a CAA-specific mouse monoclonal antibody to selectively capture virus antigen. Chicken antibodies to CAA bind to the captured antigen and are detected with horseradish peroxidase-labeled anti-chicken immunoglobulin using a conventional indirect ELISA protocol. When 388 chicken sera from specific-pathogen-free and commercial flocks from the United Kingdom, West Germany, the United States and Australia were examined, 98.5% agreement was obtained between the results of the ELISA and the indirect immunofluorescence assay. This ELISA should have worldwide application in testing SPF and commercial chicken flocks for CAA antibodies. 相似文献
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Eight-week-old shortleaf pine seedlings (Pinus echinata Mill.) with and without ectomycorrhizae formed by Pisolithus tinctorius were treated for two to eight weeks with 25 microg borate ml(-1) solution applied either to the soil, or as a foliar spray, or in both ways. Control seedlings were fertilized only with modified Hoagland's solution containing 0.03 microg ml(-1) borate. Five sugars (pinitol, fructose, glucose, myoinositol and sucrose) were quantitated in both mycorrhizal and nonmycorrhizal roots by gas-liquid chromatography. Fertilization with boron increased the total carbohydrate content of mycorrhizal roots except in seedlings receiving foliar applications of boron. Foliar + soil fertilization yielded a 24% increase in total carbohydrates in mycorrhizal roots, whereas foliar fertilization alone decreased the total carbohydrate content. Carbohydrate content of nonmycorrhizal roots was significantly increased only by soil fertilization with boron. Individual sugars were affected less by boron fertilization in nonmycorrhizal roots than in ectomycorrhizal roots. However, significant increases in sugars in response to boron fertilization were observed in both ectomycorrhizal and nonmycorrhizal plants. 相似文献
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McGiven J Hendry L Brown D Stack J Perrett L Mawhinney I 《Research in veterinary science》2008,84(1):38-40
The brucellosis surveillance scheme in Great Britain includes the serological testing of approximately 1 million bovine samples per year. These are screened by iELISA, positives going forward for confirmatory testing by CFT and SAT. Samples positive by confirmatory testing prompt substantial field investigations and interventions, but the animals involved are usually uninfected. Described below are a series of modifications to the screening method, which have resulted in a 10-fold reduction in false positive results whilst maintaining sensitivity. The key modifications include the introduction of blocking agents, a change in serum test dilution and the introduction of a control that directly defines the positive/negative cut-off. These simple modifications have had a large impact in reducing the cost of the surveillance programme due to reductions in confirmatory test requirements and a knock on effect of reducing costly field intervention. 相似文献
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D A Graham K A Mawhinney A German J C Foster B M Adair M Merza 《Journal of veterinary diagnostic investigation》1999,11(2):127-133
Isotype- and subclass-specific indirect enzyme-linked immunosorbent assays were developed to detect parainfluenza-3 virus-specific IgG1, IgG2, IgM, and IgA responses. Sera were treated with protein G-agarose prior to testing for specific IgM and IgA to eliminate the possibility of false-positive results due to IgM-rheumatoid factor and to remove interisotypic competition due to specific IgG. IgM and IgA absorbance values were expressed as a percentage of the absorbance values of positive reference sera included on each plate (S/P%), and respective positive/negative threshold values of 15.0% and 28.0% were determined. The mean interval between experimental infection of 3 calves and initial detection of specific IgG1 and IgG2 responses was 8.0 and 9.3 days respectively, rising rapidly to an initial plateau 13.7 and 11.0 days postinfection (dpi). Reinfection of these calves at 30 dpi resulted in further rapid increases, with higher plateau values reached 13.0 (IgG1) and 13.7 (IgG2) days later. The mean interval between infection and the first positive IgM and IgA responses was 6.7 and 12.3 days, respectively. IgM S/P% values peaked at 13.0 dpi, with all 3 calves showing a secondary anamnestic response to reinfection, peaking 4.7 days later. The IgA response to initial infection was weak, with only 2 calves showing an obvious peak response at 15.0 dpi. A strong anamnestic IgA response to reinfection occurred in 2 calves, with a peak response 9.5 days later. Apparent biphasic and triphasic IgM and IgA responses were evident in some calves. Acute and convalescent serum samples from 80 calves involved in 17 outbreaks of respiratory disease were tested for specific IgM and IgA. Positive IgM results were detected in 15 outbreaks, with 71 sera from 44 calves testing positive. Although IgA-positive results were detected in the same 15 outbreaks, only 42 sera from 31 calves were positive. In a previous study, seroconversion was detected in 21 of these calves from 10 outbreaks. Thus the diagnostic potential of the assays was in the order IgM > IgA > seroconversion. The correlations between IgM and IgA, IgM and seroconversion, and IgA and seroconversion results for each calf were 73.8%, 58.8% and 62.5%, respectively. 相似文献
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HY Jang YH Kim BW Kim IC Park HT Cheong JT Kim CK Park HS Kong HK Lee BK Yang 《Reproduction in domestic animals》2010,45(6):943-950
Melatonin, the major secretory product of the pineal gland, scavenges a variety of reactive oxygen and nitrogen species in vivo and in vitro, indicating that melatonin is a potent function as an antioxidant. The objective of this study was to investigate the effect of melatonin in the presence or absence of hydrogen peroxide (H2O2) on sperm characteristics (motility, viability, survival rate, membrane integrity, lipid peroxidation (LPO) and mitochondria activity) and also to examine the developmental rates to the blastocysts stage of porcine oocytes fertilized in vitro with semen treated with or without melatonin (100 nm ) in the presence or absence of H2O2 (250 μm ). The sperm were treated with melatonin in the presence or absence of H2O2 for 3, 6, 9 and 12 h at 37°C and then analysed for the sperm characteristics. The porcine embryos were produced by in vitro maturation and in vitro fertilization (IVM/IVF) using semen treated with or without melatonin (100 nm ) in the presence or absence of H2O2 (250 μm ) for 6 h. The semen characteristics, including motility, viability, survival rate, membrane integrity and mitochondria activity, were higher in the groups that were treated with melatonin in comparison to other groups, irrespective of incubation periods. Malondialdehyde levels in control, melatonin and melatonin + H2O2 groups were lower than H2O2 only group. A positive correlation was shown among motility, viability, survival rate and membrane integrity, but a negative correlation was observed between LPO and the other evaluation methods. The developmental rates to blastocysts of IVM/IVF porcine oocytes fertilized by semen treated with melatonin were significantly increased compared with any other groups, with the cell number of blastocysts shown to have a similar trend to the developmental rates. These results demonstrate that melatonin can improve the semen characteristics during in vitro storage and support the developmental ability of IVM/IVF embryos in pigs. 相似文献
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HY Jang SJ Ji YH Kim HY Lee JS Shin HT Cheong JT Kim IC Park HS Kong CK Park BK Yang 《Reproduction in domestic animals》2010,45(6):967-974
The aim of the present study was to elucidate the fundamental mechanism of bovine oviduct epithelial cell (BOEC) co‐culture on developmental capacity of bovine in vitro oocyte maturation/in vitro fertilization (IVM/IVF) embryos. We examined the effects of astaxanthin against nitric oxide‐induced oxidative stress on cell viability by MTT assay, lipid peroxidation (LPO) by using thiobarbituric acid (TBA) reaction for malondialdehyde (MDA) and the expression of antioxidant genes (CuZnSOD, MnSOD and Catalase) or apoptosis genes (Bcl‐2, Caspase‐3 and Bax) by RT‐PCR in BOEC. We also evaluated the developmental rates of bovine IVM/IVF embryos co‐cultured with BOEC pre‐treated with astaxanthin (500 μm ) in the presence or absence of sodium nitroprusside (SNP, 1000 μm ) for 24 h. Cell viability in BOEC treated with SNP (50–2000 μm ) lowered, while astaxanthin addition (50–500 μm ) increased it in a dose‐dependent manner. Cell viability in astaxanthin plus SNP (1000 μm ) gradually recovered according to the increase in astaxanthin additions (100–500 mm ). The LPO in astaxanthin group (50–500 μM) gradually decreased in a dose dependent manner and among SNP or astaxanthin plus SNP group, SNP alone and astaxanthin (50 μM) plus SNP shown a significant increase than other groups (p < 0.05). Expression of apoptosis or antioxidant genes was detected by RT‐PCR. Bcl‐2 and antioxidant genes were detected in astaxanthin or astaxanthin plus SNP group, and Caspase‐3 and Bax genes were only found in SNP group. When bovine IVM/IVF embryos were cultured for 6–7 days under co‐culture system such as BOEC treated with astaxanthin in the presence or absence of SNP, the developmental ability to blastocysts in 500 μm astaxanthin group was the highest of all groups. These results suggest that astaxanthin has a antioxidative effect on cell viability and LPO of BOEC, and development of bovine IVM/IVF embryos due to the induction of antioxidant genes and suppression of apoptosis genes. 相似文献
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There are several skin grafting methods described in the human and animal literature. Currently, there are five types of free grafts used in horses: pinch and punch grafts, split and full-thickness sheet or mesh grafts and tunnel grafts. Published methods of tunnel grafting describe the use of alligator forceps. The alligator forceps create a poor tunnel and are excessively traumatic to the granulation bed. This technique utilised a 13G Jamshidi needle that was placed across the granulation bed and created a uniform tunnel. The Jamshidi needle was atraumatic to the granulation bed increasing the opportunity for graft survival. A twin bladed scalpel allowed for the quick creation of uniform width grafts. Removal of the overlying tunnel ‘roof’ took place 5–14 days later to allow graft expansion. This case series included five horses with distal limb wounds and one with a wither injury. Four horses required general anaesthesia for graft placement and three required general anaesthesia for the removal of the tunnel roof. The acceptance of the grafts varied from 70% to 100%. Graft expansion to cover the granulation tissue took 2–5 months. This case series demonstrates that this technique of graft production and placement is an easy method for achieving successful skin grafting. Compared to other graft types, tunnel grafts are more readily accepted. Cosmetic and functional results achieved are better than those with pinch and punch grafts. Tunnel grafting does not require expensive equipment or advanced training, and in some cases can be performed under standing sedation. 相似文献
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Production and preliminary characterization of monoclonal antibodies to chicken anemia agent 总被引：2，自引：0，他引：2
M S McNulty D P Mackie D A Pollock J McNair D Todd K A Mawhinney T J Connor F McNeilly 《Avian diseases》1990,34(2):352-358
Mice were immunized with partially purified preparations of the Cux-1 isolate of chicken anemia agent (CAA), and their splenocytes were fused with NSO myeloma cells. Three patterns of staining of CAA-infected cells were recognized when the resulting hybridomas were screened by indirect immunofluorescence (IIF). Hybridomas representative of each staining pattern were cloned, and the monoclonal antibodies (MAbs) were characterized. Type 1 staining was indistinguishable from that produced by polyclonal chicken antisera to CAA. Type 2 staining was confined to large nuclear inclusions. Type 3 staining was predominantly nuclear and granular, and differed from type 1 in being more intense and occurring in a higher proportion of nuclei. Three MAbs producing type 1 staining were predominantly Cux-1-specific by IIF; they also reacted to lower titers with the Gifu-1 isolate but not at all with three other CAA isolates. These MAbs had very slight neutralizing activity against Cux-1. Another MAb giving type 1 staining reacted with all CAA isolates tested to high titers in IIF and neutralization tests. MAbs with type 2 and type 3 staining reacted by IIF with all CAA isolates tested but possessed no neutralizing activity. The availability of MABs to CAA should facilitate development of diagnostic tests for the virus. 相似文献