Application of an indirect carbon immunoassay (CIA) for the rapid diagnosis of antibody to Toxoplasma gondii in sheep

Carbon immunoassay (CIA), a novel indirect rapid test for Toxoplasma antibody in sheep, was compared with indirect fluorescent antibody assay (IFA). CIA relies on the adherence of carbon particles of India-ink to rabbit immunoglobulin G.l Carbon labelled anti-sheep rabbit IgG was used for the detection of sheep antibody when attached to tachyzoites of Toxoplasma gondii. The result was read in an ordinary light microscope and there was a clearcut difference between negative and positive reactions. Out of a total of 97 sheep sera tested, 15 sera were negative in both tests and 12 were negative in CIA but showed low positive titres in IFA. The remaining 70 sera were positive in both tests but the titres were usually about 3 dilution steps lower when investigated with CIA as compared to IFA.     

鸡传染性贫血病疫苗免疫母鸡后子代雏鸡外周血液T细胞数量及免疫球蛋白含量变化

应用免疫SPA菌体不法和间接ELISE法对鸡传染性贫血病（CIA）疫苗免疫母鸡后，其子代雏鸡外周血液T细胞数量和血清IgG,IgM,IgA含量进行了动态研究。结果发现：CIA疫苗免疫母鸡后，其子代雏鸡外周血液T细胞数量和IgG,IgM,IgA含量均不同和蔼地高于未免疫的相对照雏鸡，表明CIA疫苗免疫母后，其子代雏鸡全身的体液上述各项指标均明显低于疫苗免疫的子代雏鸡，这与CIAV强毒攻击子代雏鸡后，未免疫的子代雏鸡外周血液上述各项指标均明显低于疫苗免疫的子代雏鸡，这与CIAV强毒感染雏鸡后，其免疫器官组织，尤其是胸腺严重损害，淋巴细胞变性坏死子等密切相关，是导致感染鸡免疫功能低下的基础。     

Prediction of crude fat content of longissimus muscle of beef using the ratio of fat area calculated from computer image analysis: comparison of regression equations for prediction using different input devices at different stations

Crude fat content of longissimus (ribeye) muscle of beef cattle was predicted from a ratio of fat area (RFA) to area of ribeye muscle calculated from computer image analysis (CIA). Cross sections of 64 ribeyes taken from the 6-7th rib from cattle at experiment station A and cross sections of 94 ribeyes taken from the 6-7th rib from cattle at Experiment Station B were used in this study. Slices (1 to 1.5 cm in thickness) of just the Longissimus dorsi were homogenized and sampled for chemical estimation of crude fat content using petroleum ether. Crude fat content as determined from chemical analysis was used as the true estimate of fat content. A CCD (charge-coupled device) camera was used as the input device at Experiment Station A, and a single-lens reflex camera was used at Experiment Station B to photograph ribeyes for CIA. The contour comparison method, which assigns a threshold value for each marbling particle, was used to obtain accurate binarization in this study. Minimum and maximum of chemical measurements of crude fat were 2.1 and 39.8%, and for CIA calculation of the RFA were 6.1 and 56.8%, respectively. This range covered almost the complete range of the beef marbling standard used in carcass grading in Japan. The equation for the regression of the crude fat content (Y) on RFA (X) calculated from CIA for all of the data was Y = .793X-3.04 with r2 = .96. Regression equations for prediction of crude fat percentage from RFA taking into consideration the effect of experiment station were Y = .741X-2.22 with r2 = .91 for Experiment Station A, and Y = .782X-2.54 with r2 = .91 for Experiment Station B. Analysis of covariance showed that the effects of experiment stations on intercepts and slopes were not significant (P > .10). The ranges of differences between actual and predicted crude fat content from the prediction equation that was calculated without consideration of the effect of station were -6.4 to 4.0%. CIA of cross sections of the ribeye muscle seems to have potential for prediction of crude fat content.     

CIA免疫母鸡后子代雏鸡免疫器官T细胞变化的研究

鸡传染性贫血病（CIA）疫苗免疫母鸡后，对其子雏鸡免疫器官的T细胞变化进行了动态研究。结果发现，CIA疫苗免疫母鸡后，子代雏鸡免疫器官T细胞数量较未免疫对照子代雏鸡明显增加；强毒攻击子代雏鸡后，免疫攻毒组雏鸡免疫器官T细胞数量在13日龄内明显高于未免疫攻毒组雏鸡。表明CIA疫苗可使子代雏鸡免疫器官的细胞免疫功能增强，能抵卸强毒攻击。     

鸡传染性贫血病免疫母鸡后子代雏鸡免疫器官抗体生成细胞变化

鸡传染性贫血病疫苗免疫母鸡后，对其于代雏鸡免疫器官的抗体生成细胞变化进行了动态研究。结果发现，CIA疫苗免疫母鸡后，于代雏鸡免疫器官抗体生成细胞数量较未免疫对照于代雏鸡明显增加；强毒攻击于代雏鸡后，免疫攻毒组雏鸡免疫器官抗体生成细胞数量在27日龄内明显高于未免疫攻毒组雏鸡。表明CIA疫苗可使于代雏鸡免疫器官的体液免疫功能增强，能抵御强毒攻击。     

Humoral antibody response of rabbits to experimental infection with Encephalitozoon cuniculi

Six female rabbits (Oryctolagus cuniculus) were experimentally infected intravenously with approximately 1.5 X 10(7) live spores of Encephalitozoon cuniculi. Head tilt was observed as the single clinical sign in only one of the six animals. Antibody response was registered over 68 days postinfection using the indirect immunofluorescence test (IFT) for IgM and IgG, and the carbon immunoassay (CIA). IgG titers reached a level of 160-2560 after a latent phase of 13-28 days, followed by a 2-4 week relatively steep increase. The IgM seroconversion was faster than that of IgG and occurred at the beginning of the antibody response. Thus, simultaneous detection of both IgM and IgG allowed the infection to be identified as recent. Long, short, and episodic antibody responses could be distinguished: the IgG titer continued to increase on Day 68 in one animal (long response) and began to decrease between Days 45 and 63 in three other animals (short response). In two additional animals the seroconversion was very short, occurring between Days 13 and 41, and 28 and 52, respectively (episodic response). The CIA proved to be specific, reliable, and simple to perform; titers were slightly higher than in the IFT. Parasite pseudocysts were detected scattered throughout the brain on Day 68 in four of the six rabbits. The persistence of antigen in the brain did not correlate with antibody response, which in most cases was shorter.     

Chicken infectious anemia in Mexico: virus identification and serology survey.

Three chicken infectious anemia (CIA) virus strains were isolated from 10 different sick broiler and replacement chicken flocks with the MDCC-MSB1 cell line. One-day-old specific-pathogen-free chicks were inoculated later, with the three original samples being positive in tissue culture; one induced signs and lesions, another only lesions typical for CIA. One isolate was selected for further trials and showed resistance to chloroform and heat (75 C for 5 min) and passed through a 45-nm filter membrane but did not pass through the 22-nm filter. These characteristics were similar to the Del Rose reference strain of chicken anemia virus. By electron microscopy, the diameter of particles obtained from the pellet of infected cell cultures was between 22 and 27 nm. Serology survey carried out with 580 serum samples from different poultry farms all over the country with a commercial enzyme-linked immunosorbent assay kit gave proof of widespread seroconversion, indicating that CIA should be considered endemic to Mexico.     

鸡传染性贫血病毒的分子生物学及其核酸检测技术

鸡传染性贫血 (CIA)是鸡的重要的免疫抑制性疾病之一 ,以引起雏鸡再生障碍性贫血和全身淋巴组织萎缩为主要特征。鸡传染性贫血病毒 (CIAV)主要引起鸡骨髓成红细胞和胸腺皮质的成淋巴细胞溶细胞感染 ,继而导致贫血和免疫抑制。该病毒在世界范围内广泛存在 ,是养鸡业潜藏的巨大威胁。文章从 CIA的危害、基本控制方法、病原的基因组及其所编码的蛋白质的功能、疾病的诊断等方面的最新进展进行了综述 ,重点介绍了病原的分子生物学研究进展及其核酸检测技术 ,并对 CIAV的研究前景进行了展望     

鸡传染性贫血疫苗免疫母鸡后子代雏鸡的免疫学变化

应用免疫学新技术对鸡传染性贫血（CIA）疫苗免疫母鸡后，其子代雏鸡外周血液和免疫器官组织及局部体液的免疫学变化进行了动态研究。结果表明：CIA疫苗免疫母鸡后，其子代雏鸡外周血液T、B细胞数量和IgG、IgM、IgA含量及免疫器官组织的T细胞和IgG、IgM、IgA抗体生成细胞以及泪液、气管液、胆汁、肠液的IgA、IgM、IgG含量均不同程度地高于未免疫的相应对照雏鸡，表明CIA疫苗免疫母鸡后，其子代雏鸡全身的体液免疫和细胞免疫功能明显增强。而CIAV强毒攻击后，未免疫的子代雏鸡，其外周血液和免疫器官组织及局部体液的上述各项指标均明显低于疫苗免疫的子代雏鸡。     

鸡贫血病的最新研究进展

鸡贫血病是由鸡贫血病毒（Chicken anemia virus,CAV)引起的鸡传染性贫血。本病于1979年由日本学者Yuasa等首先报道，并分离到病毒，其后英国、瑞典、德国、加拿大、美国及中国等国家都证实了本病的存在，澳大利亚及其它一些欧洲、非洲、亚洲国家，通过血清学调查，也发现了鸡群广泛感染CAV^[1]。现在已用PCR、核酸探针等多种方法快速诊断该病^[2]。目前，鸡贫血病已被当作重要的世界性禽病。本文就近几年来，在该病的流行病学、致病机理、病原特征、诊断技术及防治方法方面的最新进展作一介绍。     

Economically Important Non-oncogenic Immunosuppressive Viral Diseases of Chicken—Current Status

Immunosuppressive viral diseases threaten the poultry industry by causing heavy mortality and economic loss of production, often as a result of the chickens' increased susceptibility to secondary infections and sub-optimal response to vaccinations. This paper aimed to present an up-to-date review of three specific economically important non-oncogenic immunosuppressive viral diseases of chickens, viz. chicken infectious anaemia (CIA), infectious bursal disease (IBD) and hydropericardium syndrome (HPS), with emphasis on their immunosuppressive effects. CIA and IBD causes immunosuppression in chickens and the socio-economic significance of these diseases is considerable worldwide. CIA occurs following transovarian transmission of chicken anaemia virus and has potential for inducing immunosuppression alone or in combination with other infectious agents, and is characterized by generalized lymphoid atrophy, increased mortality and severe anemia. The virus replicates in erythroid and lymphoid progenitor cells, causing inapparent, sub-clinical infections that lead to depletion of these cells with consequent immunosuppressive effects. The IBD virus replicates extensively in IgM(+) cells of the bursa and chickens may die during the acute phase of the disease, although IBD virus-induced mortality is highly variable and depends, among other factors, upon the virulence of the virus strain. The sub-clinical form is more common than clinical IBD because of regular vaccination on breeding farms. Infection at an early age significantly compromises the humoral and local immune responses of chickens because of the direct effect of B cells or their precursors. HPS is a recently emerged immunosuppressive disease of 3-6-weeked broilers, characterized by sudden onset, high mortality, typical hydropericardium and enlarged mottled and friable livers, with intranuclear inclusion bodies in the hepatocytes. The agent, fowl adenovirus-4, causes immunosuppression by damaging lymphoid tissues; the presence of IBD and CIA viruses may predispose for HPS or HPS may predispose for other viral infections. Synergism with CIA or other virus infections or prior immunosuppression is necessary to produce IBH-HPS in chickens and the susceptibility of chickens infected with fowl adenovirus varies throughout the course of CIA infection. The mechanism of immunosuppression has been studied in detail for certain chicken viruses at molecular levels, which will provides new opportunities to control these diseases by vaccination.     

An indirect ELISA for detection of Encephalitozoon cuniculi infection in farmed blue foxes (Alopex lagopus)

Infection with the intracellular microsporidium Encephalitozoon cuniculi can cause serious disease, encephalitozoonosis, in the blue fox (Alopex lagopus). The disease diagnosis is based on clinical signs and pathological findings, and detection of E. cuniculi or circulating antibodies directed against the parasite. Indirect immunofluorescence (IFAT) and carbon immunoassay (CIA) are the most commonly used serological methods for diagnosis in this species. In the present study, an indirect ELISA (enzyme linked immunosorbent assay) was established and evaluated against IFAT by testing of 205 field samples from blue foxes. There was high agreement between the results of the ELISA and CIA (kappa=0.99), and the ELISA and IFAT (kappa=0.958). There was no significant statistical difference between the tests (p>0.05). It was concluded that the ELISA could be used to identify seropositive farmed blue foxes. The advantage of the ELISA lies in the potential of screening large numbers of animals with the goal of eradicating E. cuniculi infection in the farms.     

鸡传染性贫血免疫对肝微粒体抗氧化酶活性的影响

目的是观察鸡传染性贫血多次免疫对肝微粒体中抗氧化酶活性的影响.对20只SPF鸡随机分为2组,每组10只,免疫组鸡用鸡传染性贫血弱毒苗免疫4次,每次间隔2周,对照组鸡注射同剂量的生理盐水.最后一次免疫后10d取肝脏制备微粒体,利用测试盒测定肝微粒体中的GSH-Px活性、SOD活性、CAT活性和MDA含量.结果与对照组相比,免疫组肝微粒体中GSH Px活性、SOD活性和CAT活性都显著提高(P＜0.05),MDA含量显著减低(P＜0.05).结论为鸡传染性贫血多次免疫可提高鸡体抗氧化能力.     

鸡贫血病-法氏囊病联合免疫母鸡后子代雏鸡外周血液T细胞数量及免疫球蛋白含量变化

应用免疫SRA菌体花环法和间接ELISE法对鸡传染性贫血病 (CIA) -传染性法氏囊病 (IBD)联合免疫母鸡后 ,其子代雏鸡外周血液T细胞数量和免疫球蛋白IgG、IgM、IgA含量变化进行了动态研究。结果发现 ,CM -IBD联合免疫母鸡后 ,其子代雏鸡外周血液T细胞数量和IgG、IgM、IgA含量均不同程度地高于未免疫的相应对照雏鸡 ,表明CIA-IBD联合免疫母鸡后 ,其子代雏鸡外周血液体液免疫和细胞免疫功能明显增强。而CIAV、IBDY强毒攻击后 ,未免疫的子代雏鸡 ,其外周血液的上述指标均明显低于疫苗免疫的子代雏鸡 ,表明未免疫的子代雏鸡外周血液的免疫机能降低 ,这与未免疫雏鸡缺乏特异性抗体 ,强毒攻击后 ,雏鸡免疫器官组织广泛损害 ,淋巴细胞变性坏死等有关     

Micro-computed tomography evaluation and pathological analyses of female rats with collagen-induced arthritis

Imaging techniques have been introduced to assess the efficacy and toxicity of developing pharmaceuticals. The purpose of this study was to perform a comprehensive characterization of collagen-induced arthritis (CIA) in rats using micro-computed tomography (micro-CT) and to compare the results with data from conventional pathological examination. Arthritis was induced by collagen in 24 female Wistar rats. Micro-CT and pathological analyses were performed to assess arthritis progression. Micro-CT analysis showed marked joint destruction occurring in a time-dependent manner following collagen administration. Bone volume was significantly decreased in the tibia at weeks 3 and 4 compared to week 0 (p < 0.05 and p < 0.01, respectively). Additionally, percent bone volume was significantly reduced in the tibia at week 4 compared to week 0 (p < 0.05). In contrast, bone surface/bone volume and trabecular separation were significantly increased in the tibia of the animals at week 4 compared to week 0 (p < 0.05). Severe joint destruction with extensive inflammation, erosion of cartilage and bone, and infiltration of inflammatory cells were observed in the knee joints of the collagen-treated rats. Taken together, micro-CT made it possible to quantify CIA lesions and should be performed with pathological examination in rats.     

Serological survey of Encephalitozoon cuniculi in farm rabbits in Italy

Rabbit sera (n = 1600) from 40 commercial farms were submitted to a serological screening for Encephalitozoon cuniculi by an enzyme-linked immunosorbent assay (ELISA) and a carbon immunoassay (CIA test). Antibodies anti-Encephalitozoon cuniculi were found in 505/1600 (31.6%) sera analysed, and all the farms (100%) resulted positive. Rabbits older than 4 months showed a significantly higher seropositivity for E. cuniculi (chi-squared test: p < 0.0001) than rabbits under 4 months, E. cuniculi sero-prevalence showed an increasing trend in rabbits within the farm along with the increase in the “number of rabbits on the farm”; however, this trend was not significant (Spearman’s correlation: p = 0.073).The findings of the present study confirm that rabbit is the main reservoir of E. cuniculi; they are of epidemiological relevance and immediate public health importance because of the recognized infectivity in humans by the microsporidium.     

Serological survey for antibodies to Encephalitozoon cuniculi in rabbits in Taiwan

Encephalitozoon cuniculi (E. cuniculi) is a microsporidian parasite commonly found in rabbits that can infect humans, causing encephalitozoonosis. Our laboratory recently confirmed the first case of encephalitozoonosis in a rabbit in Taiwan; the prevalence of encephalitozoonosis is not well documented, even when many clinics suspect pet rabbits as being infected. This study surveys the seropositivity of E. cuniculi using carbon immunoassay (CIA) and enzyme-linked immunosorbent assay (ELISA). Serological examination of 171 rabbits using CIA and ELISA showed that 63.2% (108/171) and 67.8% (116/171) were seropositive against E. cuniculi, respectively. Thirteen of the 14 rabbits (92.9%) with neurological symptoms were seropositive. Except for gender, health status and location had a significant effect on E. cuniculi seropositivity (p<0.05). Adult rabbits aged older than 4 months exhibited significantly higher seropositivity for E. cuniculi than young rabbits (p<0.05). In conclusion, this study shows that E. cuniculi is present and widespread among healthy rabbits in Taiwan. Therefore, the fields of veterinary and human medicine in Taiwan should be aware of this zoonotic issue and the resulting public health concern of encephalitozoonosis.     

Computer image analysis traits of cross-sectioned dry-cured hams: a genetic analysis

The aims of this study were to estimate genetic parameters of image analysis traits of cross-sectioned dry-cured hams and carcass weight (CW) and to investigate effects of some nongenetic sources of variation on these traits. Computer image analysis (CIA) had been carried out for digital images of the cross-section of 1,319 San Daniele dry-cured hams. The cross-sectional area (SA, cm(2)); the average thickness of subcutaneous fat (FT, cm); and the proportions of lean (LA, %), fat-eye (FEA, %), and subcutaneous fat area (SCF, %) to SA, and of biceps femoris (BFA, %) and semitendinosus muscle area (STA, %) to LA were recorded. Bivariate analyses were carried out for pairs of traits for estimation of genetic parameters using Bayesian methodology and linear models. Linear models included the nongenetic effects of slaughter groups and sex and the additive genetic effects of pigs and their ancestors (1,888 animals). Variation of FEA was nearly 4-fold that of SA and LA. Variation of CIA traits due to sex effect was not large, whereas slaughter group effects were relevant sources of variation for all traits. For all traits, with the exception of FEA, the posterior probability for the true heritability being greater than 0.1, was greater than 0.95. Point estimates of heritabilities for FT and SCF were 0.42 and 0.51, respectively. Heritability estimates for FEA, LA, BFA, and STA were 0.13, 0.44, 0.44, and 0.36, respectively. The genetic correlations between CW and CIA traits were positive and large for SA (0.86), positive and moderate for FT, FEA, and STA (0.47, 0.40, and 0.45, respectively) and negative with LA (-0.28). Although FEA, FT, and SCF were all measures of the extent of fat deposition in the ham, the genetic correlations between FT or SCF and FEA were very low. A very large estimate (0.74) was obtained for the genetic relationship between SA and FEA, suggesting that reduction of ham roundness through selective breeding would be beneficial for decreasing FEA. On the basis of the estimated parameters, genetic selection is expected to be effective in changing size of fatty and lean areas of the cross-section of dry-cured hams. Causes related to the abnormal development of the fat-eye depot remain unknown, but this study provided evidence that influences of polygenic effects on phenotypic variation of FEA are limited.     

鸡贫血病灭活疫苗免疫后雏鸡免疫状况研究

应用鸡传染性贫血病灭活疫苗免疫雏鸡后,１０、２０、３５、４５天检测了空白对照组、免疫组的外周血液、胸腺、脾脏和法氏囊内ＣＤ３＋、ＣＤ４＋、ＣＤ８＋、γδＴ细胞亚群、ＩｇＧ阳性细胞的变化;并且在３５天、４５天检测免疫攻毒组和攻毒组上述细胞免疫和体液免疫指标的变化。结果显示,应用贫血病灭活疫苗后免疫组雏鸡的Ｔ细胞亚群的ＣＤ４／ＣＤ８比值出现明显降低,ＩｇＧ阳性细胞有升高变化;免疫攻毒组雏鸡状态明显好于攻毒组。研究表明,鸡传染性贫血病疫苗可以激发机体体液免疫反应,并在一定程度上诱导ＣＴＬ活化,对ＣＡＶ攻击产生足够的免疫保护。     

Serological survey for antibodies to Encephalitozoon cuniculi in pet rabbits in Italy

Pet rabbits (n = 125) from Southern Italy were submitted to a serological screening for Encephalitozoon cuniculi, using an enzyme-linked immunosorbent assay (ELISA) and a carbon immunoassay (CIA). Seventy-eight examined rabbits showed clinical signs suggestive of encephalitozoonosis (head tilt, ataxia, paralysis, cataracts, uveitis, polyuria and polydipsia), whereas 47 were healthy rabbits. Antibodies anti-E. cuniculi were found in 84/125 (67.2%) sera analysed. The results of the chi-squared test showed that sex and health status had no significant effect (P > 0.05) on E. cuniculi seropositivity; however, rabbits older than 4 months had a seropositivity for E. cuniculi significantly (P < 0.05) higher than that of rabbits aged up to 4 months. The results of the present survey reinforce the assumption that rabbit may be indicated as the main reservoir of E. cuniculi; therefore, routine screening examinations in pet rabbits are strongly advised considering the zoonotic potential of this parasite.