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奶作为一种新的诊断工具,用于快速检测奶山羊的 Fascioliasis,使用排泄/分泌抗原。

Milk As a New Diagnostic Tool for Rapid Detection of Fascioliasis in Dairy Goats Using Excretory/Secretory Antigen.

机构信息

Food Hygiene and Control, Faculty of Veterinary Medicine, Cairo University, P.O Box 12211, Giza, Egypt.

Parasitology Departments, Faculty of Veterinary Medicine, Cairo University, P.O Box 12211, Giza, Egypt.

出版信息

Acta Parasitol. 2021 Jun;66(2):336-345. doi: 10.1007/s11686-020-00286-z. Epub 2020 Sep 29.

Abstract

PURPOSE

Fascioliasis is a serious livestock illness of particular importance for dairy goats; the objectives of this study were to describe effects of F. gigantica on milk composition and to use this information to estimate economic damages linked with Fasciola spp. infections. Furthermore, the study sought to standardize the use of milk instead of serum for early diagnosis of fascioliasis in dairy goats.

METHODS

One-hundred samples of goat milk along with corresponding blood samples were obtained at random from flocks in Cairo and Giza Governorates. The ELISA and DOT-ELISA were performed in both serum and milk of dairy goats.

RESULTS

Total mesophilic count (mean value) was 2.12 × 10 ± 1.63 × 10 CFU/ml in enzyme-linked immunosorbent assay (ELISA) positive samples and 1.46 × 10 ± 8.58 × 10 CFU/ml in ELISA-negative samples. The mean values were significantly different (P < 0.05). The mean values of percentages of fat, SNF, protein, salts, lactose, pH, and MSCC/ml in ELISA-positive samples were 2.3 ± 0.17, 8.21 ± 0.63, 3.08 ± 0.18, 0.90 ± 0.06, 3.64 ± 0.28, 6.93 + 0.53, and 1.18 × 10 ± 9.07 × 10 cells/ml, respectively. A significant difference (P < 0.05) between the mean values of two composition parameters, i.e., percent of fat and MSCC/ml in ELISA-positive and -negative samples, for Fasciola gigantica was observed. The antigen used for the diagnosis of F. gigantica was excretory/secretory (E/S) antigen. The dilutions of (E/S) concentrations after checkerboard titration for indirect ELISA were 20 μg/ml protein and for dot-ELISA, 300 ng/μl. Sera dilution was 1:100 in the two tests, and milk dilution was 1:50 for indirect ELISA, and 1:25 for dot-ELISA. The two tests were performed using known F. gigantica positive and negative goat sera and known rat hyper immunized negative and positive sera against E/S antigen of F. gigantica as well as known sera for paramphistomes without F. gigantica infection. The cutoff values in indirect ELISA were 0.45 for sera and 0.35 for milk.

CONCLUSION

The application of different serological technique in goat farms reveals a good test in rapid diagnosis of fascioliasis especially the uses of dot ELISA when using the milk instead of the serum.

摘要

目的

片形吸虫病是一种严重的家畜疾病,对奶山羊尤为重要。本研究的目的是描述巨型片形吸虫对乳成分的影响,并利用这些信息估计与片形吸虫属感染相关的经济损失。此外,本研究旨在标准化使用牛奶代替血清进行奶山羊早期片形吸虫病诊断。

方法

从开罗省和吉萨省的羊群中随机抽取 100 份山羊奶样本和相应的血液样本。在奶山羊血清和奶中进行酶联免疫吸附试验(ELISA)和斑点酶联免疫吸附试验(DOT-ELISA)。

结果

酶联免疫吸附试验(ELISA)阳性样本的总嗜中性粒细胞计数(平均值)为 2.12×10±1.63×10 CFU/ml,ELISA 阴性样本的总嗜中性粒细胞计数(平均值)为 1.46×10±8.58×10 CFU/ml。平均值差异显著(P<0.05)。ELISA 阳性样本中脂肪、非脂乳固体、蛋白质、盐、乳糖、pH 和 MSCC/ml 的百分比的平均值分别为 2.3±0.17、8.21±0.63、3.08±0.18、0.90±0.06、3.64±0.28、6.93+0.53 和 1.18×10±9.07×10 细胞/ml。ELISA 阳性和阴性样本中两个组成参数(脂肪百分比和 MSCC/ml)的平均值之间存在显著差异(P<0.05)。用于诊断巨型片形吸虫的抗原是排泄/分泌(E/S)抗原。间接 ELISA 的棋盘滴定后 E/S 浓度的稀释度为 20μg/ml 蛋白,点 ELISA 为 300ng/μl。两种试验的血清稀释度均为 1:100,间接 ELISA 的奶稀释度为 1:50,点 ELISA 的奶稀释度为 1:25。两种试验均使用已知的巨型片形吸虫阳性和阴性山羊血清,已知的大鼠抗巨型片形吸虫 E/S 抗原高免疫阴性和阳性血清以及已知的无片形吸虫感染的副血矛线虫血清进行。间接 ELISA 的截断值为血清 0.45,奶 0.35。

结论

在山羊养殖场应用不同的血清学技术可以快速诊断片形吸虫病,特别是在使用牛奶代替血清时使用点 ELISA。

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