禽呼肠病毒琼扩抗原和血清的制备

以禽呼肠病毒S1133毒株尿囊腔接种10日龄无特定病原体(SPF)鸡胚,无菌收集24 h～120 h的死胚尿囊液,加1 mL/L的甲醛灭活,浓缩制得琼扩抗原;用该抗原制成灭活苗两次免疫SPF鸡,无菌采血分离制备阳性血清;选用SPF鸡无菌采血分离制备阴性血清.经过效价测定制备16单位琼扩抗原,8单位阳性血清;取免后的SPF鸡血样4个20～2-5稀释分别与1单位～8单位琼扩抗原做琼扩试验,确定了应该用4单位或8单位琼扩抗原做工作抗原;并以血清中和试验为参照证明琼扩试验在血清抗体监测中的可行性.本研究成功制备了禽呼肠病毒琼扩抗原与血清,为用琼扩试验定量检测禽呼肠病毒感染禽和疫苗免疫禽的抗体奠定了基础.     

An Acute-phase Protein as a Regulator of Sperm Survival in the Bovine Oviduct: Alpha 1-acid-glycoprotein Impairs Neutrophil Phagocytosis of Sperm In Vitro

We have previously shown that polymorphonuclear neutrophils (PMNs) are present in bovine oviduct fluid under physiological conditions, and that the oviduct provides a microenvironment that protects sperm from phagocytosis by PMNs. Alpha 1-acid glycoprotein (AGP) is a major acute-phase protein produced mainly in the liver that has immunomodulatory functions. AGP mRNA is expressed in extrahepatic organs, such as the lung, kidney, spleen, lymph node, uterus, and ovary. Therefore, in this study, we investigated, 1) the local production of AGP in the bovine oviduct, 2) the effect of AGP on the phagocytic activity of PMNs for sperm and superoxide production and 3) the impact of AGP desialylation on the PMN phagocytosis of sperm. The AGP gene was expressed in cultured bovine oviduct epithelial cells (BOECs) and AGP protein was detected in oviduct fluid. Preexposure of PMNs to AGP at physiological levels impaired PMN phagocytosis for sperm and superoxide generation. The desialylation of AGP eliminated these suppressive effects of AGP on PMN. Scanning electron microscopy revealed that AGP drastically reduced the formation of DNA-based neutrophil extracellular traps (NETs) for sperm entanglement. Additionally, AGP dose-dependently stimulated BOECs to produce prostaglandin E₂ (PGE₂) which has been shown to partially contribute to the regulation of sperm phagocytosis in the bovine oviduct. AGP and PGE₂ at concentrations detected in the oviducts additively suppressed sperm phagocytosis by PMNs. These results provide evidence that locally produced AGP may be involved in protecting sperm from phagocytosis by PMNs in the bovine oviduct.     

Non-specific seroreactions against Brucella abortus in ruminants in New Zealand and the presence of Yersinia enterocolitica 0:9

The level of non-specific reactions found in the brucellosis serology of ruminants in New Zealand was very low until July 1992. This changed when, in an export consignment of 1071 deer, 35% reacted in the Brucella abortus tube agglutination test with titres varying from 50 to 200 IU. The reactors were also positive in the Rose-Bengal agglutination test and most of them reacted in the complement fixation test with titres varying from 10 to 80 IU. Yersinia enterocolitica 0:9 was later isolated from one deer of this consignment. It was the first isolate of this serotype recovered in New Zealand from an animal. Shortly after, false reactors occurred more frequently than before in sera from Brucella abortus accredited free cattle herds. As the involvement of Yersinia enterocolitica 0:9 was suspected in these cases, faecal samples from reactors and in-contact animals were cultured for this organism. Yersinia enterocolitica 0:9 was isolated from nine of 19 herds showing one or more false Brucella abortus seroreactions.

Prior to 1990, Yersinia enterocolitica serotype 0:9 had not been isolated in New Zealand, despite the recovery of a number of other bio— or serotypes of the organism from humans and animals. From 1990 onward, serotype 0:9 began to be isolated from human faecal samples with increasing frequency. Since the first isolations from deer and cattle in 1992, it has now also been recovered from a cat and an alpaca and from cattle without any association with false positive Brucella abortus reactions. All serotype 0:9 isolates were of biotype 2.     

间接ELISA,HI及AGP检测鸡血清中抗AIV抗体的敏感性比较

用鸭源Ａ型流感病毒Ｈ９Ｎ？滴鼻、点眼辅以腹腔注射,人工感染３周龄ＳＰＦ来航鸡,分别用ＡＧＰ、ＨＩ试验及间接ＥＬＩＳＡ定期测定其抗ＡＩＶ血清效价。结果,ＨＩ试验及间接ＥＬＩＳＡ于攻毒后第７～７９天显示阳性;ＡＧＰ试验从第１３～７９天呈阳性。根据首次检出血清中抗ＡＩＶ抗体的时间,间接ＥＬＩＳＡ比ＡＧＰ、ＨＩ更灵敏、快速。     

Acute phase proteins in cats: Diagnostic and prognostic role,future directions,and analytical challenges

While clinical studies on acute phase proteins (APPs) have significantly increased in the last decade, and most commercial labs are now offering major APPs in their biochemical profiles, APP testing has not been widely adopted by veterinary clinical pathologists and veterinarians. Measurement of APP concentration is a useful marker for detecting the presence or absence of inflammation in cats with various diseases. APPs can also be reliably measured in different biological fluids (eg, effusions and urine) to improve their diagnostic utility. Measurement of APPs can be extremely beneficial in cats with feline infectious peritonitis (FIP) to discriminate between FIP and non-FIP cats with similar clinical presentations. Additional benefits come from multiple and sequential measurements of APPs, particularly in the assessment of therapeutic efficacy. APPs are more sensitive than WBC counts for early detection of inflammation and to demonstrate an early remission or recurrence of the diseases. Given the potential utility of APPs, more studies are warranted, with a particular focus on the applications of APPs to guide the length of antimicrobial therapies, as suggested by the antimicrobial stewardship policy. New inflammatory markers have been discovered in human medicine, with a higher specificity for distinguishing between septic versus nonseptic inflammatory diseases. It is desirable that these new markers be investigated in veterinary medicine, to further test the power of APPs in diagnostic setting.     

琼脂扩散试验检测雏鹅新型病毒性肠炎病毒及抗体的研究

本文报道１种检测雏鹅新型病毒性肠炎病毒及抗体的琼脂扩散试验。以１／１５Ｍ ｐＨ７．２ＰＢＳ配制的含０．８５％琼脂糖凝胶检测效果最好,对ＮＧＶＥＶ提纯抗原和兔抗ＮＧＶＥＶ的ＩｇＧ的最低检出量分别为０．１５ｍｇ／ｍｌ和０．０５ｍｇ／ｍｌ。１次加样和２次加样对ＮＧＶＥＶ感染死亡雏鹅的肠、肝脏、心、脾、肾、胰腺的阳性检出率分别为１００％、８０％、４０％、５０％、２０％、６０％和１００％、１００％、５６％、     

鸡致病性MDV感染AGP法检测可靠性研究

常规琼脂扩散（ＡＧＰ）试验通过羽毛囊马立克氏病病毒（ＭＤＶ）特异抗原的检出来诊断鸡致病性ＭＤＶ的感染〔１,２,６〕,因特异性强、操作简便而广泛应用。但其检出率与感染鸡群在不同时期的检测〔３〕以及火鸡疱疹病毒（ＨＶＴ）疫苗免疫状态〔４〕密切相关。此现象...     

牛病毒性腹泻病毒地方株的分离与鉴定

从河南省不同地区规模化肉牛场牛病毒性腹泻 (BVD)疑似病例中采集病料 ,将处理好的 6份病料接种MDBK细胞 ,并盲传 4代 ,得到了 2株可产生细胞病变的病毒。 2株病毒的TCID50 分别为 10 - 5.59和 10 - 5.51;琼脂扩散试验表明 ,2株病毒均能与牛病毒性腹泻病毒 (BVDV)OregonC2 4 标准阳性血清反应 ,出现沉淀线 ;中和试验表明 ,2株病毒均能被BVDV标准阳性血清中和 ,中和指数分别为 10 3.30 和 10 3.16 ;电镜观察到圆形 ,直径为 4 0～ 6 0nm ,有囊膜 ,囊膜表面有突起的病毒粒子 ,与BVDV颗粒基本一致。因此 ,确定分离的 2株病毒均为BVDV ,分别将其命名为HN - 1株和HN - 2株。     

乳房炎奶牛乳汁体细胞数与乳汁HP、SAA和AGP水平关系的研究

对8头正常奶牛和22头自发性乳房炎奶牛的混合乳汁中血清淀粉样蛋白(SAA)、结合珠蛋白(HP)、α1-酸性糖蛋白(AGP)水平和乳汁体细胞数(SCC)的关系进行了研究.结果表明:乳房炎奶牛与正常奶牛乳汁中HP水平差异显著(P<0.05),与乳汁SCC存在极显著正相关关系(P=0.000,r=0.406);乳房炎奶牛与正常奶牛乳汁中SAA和AGP的差异不明显(P>0.05),而且与乳汁SCC的相关性也均不显著,说明乳汁HP水平能反映奶牛乳腺感染的严重程度.     

An immunoturbidimetric assay for rapid quantitative measurement of feline alpha-1-acid glycoprotein in serum and peritoneal fluid

BACKGROUND: Alpha-1-acid glycoprotein (AGP) is an acute phase protein that increases in concentration in infectious and inflammatory conditions. The serum and peritoneal fluid concentrations of AGP may be useful in the diagnosis of feline infectious peritonitis (FIP), a lethal disease of cats. Currently AGP can be measured by radioimmunodiffusion (RID) assays, which are time consuming and difficult. OBJECTIVES: The objectives of this study were to develop a rapid immunoturbidimetric assay for measurement of AGP in feline serum and peritoneal fluid and to compare the results with those obtained by RID. METHODS: AGP was purified by perchloric acid precipitation and ion-exchange chromatography from a pool of peritoneal fluid obtained from cats with FIP, as determined by a panel of laboratory tests, including serum AGP concentration, albumin: globulin ratio, and total protein concentration, anti-coronavirus antibody titers, and effusion analysis. The purified AGP in a complete Freund's adjuvant and Tween 20 mixture was injected into a sheep and blood was collected at monthly intervals. Anti-AGP antiserum, as confirmed by ELISA and Western blot techniques, and a pool of peritoneal fluid from cats with FIP were used to prepare standards. Clinical samples of feline peritoneal fluid (n=55) and serum (n=59) were assayed for AGP and results from the immunoturbidimetric and RID methods were compared. RESULTS: Significant correlation (P < .001) was obtained between methods for both peritoneal fluid (R2=.9259) and serum (R2=.9448) samples. Coefficients of variation for the immunoturbidimetric method were <5%. CONCLUSIONS: This rapid immunoturbidimetric assay for measurement of feline AGP in serum and peritoneal fluid may be of value in the diagnosis of FIP and possibly other inflammatory diseases in cats.