巴基斯坦腹泻绵羊中两种隐孢子虫病检测诊断技术的流行情况、风险因素分析与评估。
The prevalence, risk factors analysis and evaluation of two diagnostic techniques for the detection of Cryptosporidium infection in diarrheic sheep from Pakistan.
机构信息
Department of Clinical Medicine and Surgery, University of Veterinary and Animal Sciences, Lahore, Pakistan.
College of Veterinary Sciences and Animal Husbandry, Abdul Wali Khan University, Mardan, Pakistan.
出版信息
PLoS One. 2022 Jul 8;17(7):e0269859. doi: 10.1371/journal.pone.0269859. eCollection 2022.
BACKGROUND
Cryptosporidium spp. is recognized as an opportunistic zoonotic parasite that infects humans as well as wild and domestic animals. This enteric protozoan is a major cause of diarrhea in humans and animals and often result in death due to severe dehydration. The present study was designed to investigate the prevalence, identification of various risk factors and evaluation of sensitivity of the two diagnostic techniques for rapid and correct detection of Cryptosporidium infection in diarrheic sheep in Pakistan.
METHODS
A total of 360 fecal samples were collected and processed for detection of Cryptosporidium infection after proper preservation. These samples were properly stained with modified Ziehl-Neelsen acid staining and then examined under simple microscope at 100x magnification for confirmation of Cryptosporidium oocysts. The same samples were again processed through simple PCR for confirmation of the Cryptosporidium spp.
RESULTS
The age wise prevalence was detected through simple microscopy and PCR. We found highest prevalence at the age of ≤1 year followed by 1-2 years of age while the lowest prevalence was recorded at the age of ≥ 2-3 years of sheep and found significant difference between different ages (P<0.05). The sex wise prevalence showed the highest prevalence in male (♂) animals detected compared to female (♀). The overall prevalence was detected 27.08% and 18.80% through PCR and simple microscopy, respectively, and significant difference between two diagnostic techniques were observed (P<0.05). Considering the seasonality, the highest prevalence was recorded through simple microscopy in autumn, summer, and spring, while the lowest in winter. These results were confirmed through PCR.
CONCLUSION
It was concluded that molecular detection is the most efficient, specific and sensitive technique for detection of Cryptosporidium infection than simple microscopy. Moreover sheep is the major potential source of infection to other wild and domestic animals including humans.
背景
隐孢子虫被认为是一种机会性的人畜共患寄生虫，可感染人类以及野生动物和家养动物。这种肠道原生动物是人类和动物腹泻的主要原因，常因严重脱水而导致死亡。本研究旨在调查巴基斯坦腹泻绵羊中隐孢子虫感染的流行情况、各种危险因素的识别以及两种诊断技术的敏感性，以快速准确地检测隐孢子虫感染。
方法
采集并保存了 360 份粪便样本，用于检测隐孢子虫感染。这些样本经过改良的齐尔-尼尔森酸性染色适当染色后，在简单显微镜下用 100x 放大倍数检查，以确认隐孢子虫卵囊。同样的样本再次通过简单 PCR 处理，以确认隐孢子虫属。
结果
通过简单显微镜和 PCR 检测到年龄相关的患病率。我们发现，年龄在≤1 岁的羊中患病率最高，其次是 1-2 岁的羊，而≥2-3 岁的羊患病率最低，不同年龄之间存在显著差异（P<0.05）。性别相关的患病率显示，♂动物的患病率高于♀动物。通过 PCR 和简单显微镜分别检测到的总患病率为 27.08%和 18.80%，两种诊断技术之间存在显著差异（P<0.05）。考虑到季节性，秋季、夏季和春季通过简单显微镜检测到的患病率最高，冬季最低。这些结果通过 PCR 得到了证实。
结论
与简单显微镜相比，分子检测是检测隐孢子虫感染最有效、最特异和最敏感的技术。此外，绵羊是其他野生动物和家养动物（包括人类）感染的主要潜在来源。
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