Bortoluzzi C, Paras K L, Applegate T J, Verocai G G
Department of Poultry Science, College of Agriculture and Environmental Sciences, University of Georgia, Athens, GA, 30602, USA.
Department of Infectious Diseases, College of Veterinary Medicine, University of Georgia, Athens, GA, 30602, USA.
Vet Parasitol. 2018 Apr 30;254:21-25. doi: 10.1016/j.vetpar.2018.02.039. Epub 2018 Mar 6.
Monitoring Eimeria shedding has become more important due to the recent restrictions to the use of antibiotics within the poultry industry. Therefore, there is a need for the implementation of more precise and accurate quantitative diagnostic techniques. The objective of this study was to compare the precision and accuracy between the Mini-FLOTAC and the McMaster techniques for quantitative diagnosis of Eimeria maxima oocyst in poultry. Twelve pools of excreta samples of broiler chickens experimentally infected with E. maxima were analyzed for the comparison between Mini-FLOTAC and McMaster technique using, the detection limits (dl) of 23 and 25, respectively. Additionally, six excreta samples were used to compare the precision of different dl (5, 10, 23, and 46) using the Mini-FLOTAC technique. For precision comparisons, five technical replicates of each sample (five replicate slides on one excreta slurry) were read for calculating the mean oocyst per gram of excreta (OPG) count, standard deviation (SD), coefficient of variation (CV), and precision of both aforementioned comparisons. To compare accuracy between the methods (McMaster, and Mini-FLOTAC dl 5 and 23), excreta from uninfected chickens was spiked with 100, 500, 1,000, 5,000, or 10,000 OPG; additional samples remained unspiked (negative control). For each spiking level, three samples were read in triplicate, totaling nine reads per spiking level per technique. Data were transformed using log10 to obtain normality and homogeneity of variances. A significant correlation (R = 0.74; p = 0.006) was observed between the mean OPG of the McMaster dl 25 and the Mini-FLOTAC dl 23. Mean OPG, CV, SD, and precision were not statistically different between the McMaster dl 25 and Mini-FLOTAC dl 23. Despite the absence of statistical difference (p > 0.05), Mini-FLOTAC dl 5 showed a numerically lower SD and CV than Mini-FLOTAC dl 23. The Pearson correlation coefficient revealed significant and positive correlation among the four dl (p ≤ 0.05). In the accuracy study, it was observed that the Mini-FLOTAC dl 5 and 23 were more accurate than the McMaster for 100 OPG, and the Mini-FLOTAC dl 23 had the highest accuracy for 500 OPG. The McMaster and Mini-FLOTAC dl 23 techniques were more accurate than the Mini-FLOTAC dl 5 for 5,000 OPG, and both dl of the Mini-FLOTAC were less accurate for 10,000 OPG counts than the McMaster technique. However, the overall accuracy of the Mini-FLOTAC dl 23 was higher than the McMaster and Mini-FLOTAC dl 5 techniques.
由于近期家禽行业抗生素使用受到限制,监测艾美耳球虫排泄变得愈发重要。因此,需要实施更精确、准确的定量诊断技术。本研究的目的是比较Mini-FLOTAC技术和麦克马斯特技术在定量诊断家禽巨型艾美耳球虫卵囊方面的精密度和准确性。对12组经实验感染巨型艾美耳球虫的肉鸡粪便样本进行分析,以比较Mini-FLOTAC技术和麦克马斯特技术,其检测限分别为23和25。此外,使用6份粪便样本比较Mini-FLOTAC技术在不同检测限(5、10、23和46)下的精密度。为进行精密度比较,对每个样本进行5次技术重复(一份粪便悬液制作5张重复玻片),以计算每克粪便中的平均卵囊数(OPG)、标准差(SD)、变异系数(CV)以及上述两种比较方法的精密度。为比较两种方法(麦克马斯特技术、Mini-FLOTAC技术检测限5和23)之间的准确性,向未感染鸡的粪便中添加100、500、1000、5000或10000个OPG;另外的样本不添加(阴性对照)。对于每个添加水平,对3个样本进行3次重复读数,每种技术每个添加水平共读取9次。数据采用log10转换以获得方差的正态性和齐性。观察到麦克马斯特技术检测限25的平均OPG与Mini-FLOTAC技术检测限23之间存在显著相关性(R = 0.74;p = 0.006)。麦克马斯特技术检测限25和Mini-FLOTAC技术检测限23之间的平均OPG、CV、SD和精密度在统计学上无差异。尽管无统计学差异(p>0.05),但Mini-FLOTAC技术检测限5的SD和CV在数值上低于Mini-FLOTAC技术检测限23。皮尔逊相关系数显示4个检测限之间存在显著正相关(p≤0.05)。在准确性研究中,观察到对于100个OPG,Mini-FLOTAC技术检测限5和23比麦克马斯特技术更准确;对于500个OPG,Mini-FLOTAC技术检测限23的准确性最高。对于5000个OPG,麦克马斯特技术和Mini-FLOTAC技术检测限23比Mini-FLOTAC技术检测限5更准确;对于10000个OPG计数,Mini-FLOTAC技术的两个检测限都不如麦克马斯特技术准确。然而,Mini-FLOTAC技术检测限23的总体准确性高于麦克马斯特技术和Mini-FLOTAC技术检测限5。