Aboelsoued Dina, Toaleb Nagwa I, Abdel Megeed Kadria N, Hassan Soad E, Ibrahim Sally
1Department of Parasitology and Animal Diseases, National Research Centre, El Buhouth St, Dokki, Cairo, Egypt.
2Department of Animal Reproduction and AI, National Research Centre, El Buhouth St, Dokki, Cairo, Egypt.
J Parasit Dis. 2019 Sep;43(3):393-401. doi: 10.1007/s12639-019-01103-9. Epub 2019 Mar 18.
is an apicomplexan parasite of human and animals and is considered as an important co-factor in neonatal diarrhea. In this study, an explant culture was used as an in vitro model of buffalo intestine to evaluate the effect of Moringa leaves extract on () oocysts using light and scanning electron microscopy and measuring IFN-γ, IL-12 and IL-14 in the culture supernatants. oocysts were collected from naturally-infected calf feces, isolated, excysted and then co-inoculated with ileal tissue explants culture medium. The prepared Moringa leaves extract was then introduced to the infected tissues in the concentrations of 100 mg/ml and 300 mg/ml. After 24 h, tissues were collected and processed for light and scanning electron microscopy. Also, culture supernatants were collected for cytokines measurement. parasitophorous vacuoles were found attached to the surface of tissue in -infected ileal tissue explants. High magnification imaging of ileal tissue explants using scanning electron microscopy showed that Moringa leaves extracts had a great effect on -infected ileal tissue explants. There was a high significant (< 0.001) increase in IFN-γ, IL-12 and IL-14 (375, 275 and 90 pg/ml, respectively) in the supernatants of infected non-treated ileal tissue explant culture plate wells compared to the control non-infected ones (74.66, 75 and 50 pg/ml, respectively). A concentration of 100 mg/ml Moringa extract exhibited the highest anticryptosporidial effect causing a significant decrease in IFN-γ, IL-12 and IL-14 levels (225, 150 and 65 pg/ml, respectively) compared with supernatants of infected non-treated ileal explant culture plate wells. In this study, explant culturing of buffalo ileal tissues allowed investigating the pathogenesis of cryptosporidiosis using light and scanning electron microscopy and studying changes in cytokine levels in tissues with and without Moringa leaves extract treatment. This model could help to understand the regulation of intestinal secretory and inflammatory responses, and could be useful for the screening of potential anticryptosporidial candidate compounds.
是人和动物的一种顶复门寄生虫,被认为是新生儿腹泻的一个重要辅助因素。在本研究中,采用外植体培养作为水牛肠道的体外模型,利用光学显微镜和扫描电子显微镜,并通过检测培养上清液中的干扰素-γ(IFN-γ)、白细胞介素-12(IL-12)和白细胞介素-14(IL-14),来评估辣木叶提取物对()卵囊的作用。卵囊从自然感染的小牛粪便中收集,进行分离、脱囊,然后与回肠组织外植体培养基共同接种。然后将制备好的辣木叶提取物以100毫克/毫升和300毫克/毫升的浓度加入到感染的组织中。24小时后,收集组织并进行光学显微镜和扫描电子显微镜检查。同时,收集培养上清液用于细胞因子检测。在感染隐孢子虫的回肠组织外植体中发现有寄生泡附着在组织表面。使用扫描电子显微镜对回肠组织外植体进行高倍成像显示,辣木叶提取物对感染隐孢子虫的回肠组织外植体有很大影响。与未感染的对照外植体(分别为74.66、75和50皮克/毫升)相比,未处理的感染隐孢子虫的回肠组织外植体培养板孔上清液中的IFN-γ、IL-12和IL-14(分别为375、275和90皮克/毫升)有极显著升高(<0.001)。与未处理的感染隐孢子虫的回肠外植体培养板孔上清液相比,100毫克/毫升的辣木提取物浓度表现出最高的抗隐孢子虫作用,使IFN-γ、IL-12和IL-14水平显著降低(分别为225、150和65皮克/毫升)。在本研究中,水牛回肠组织的外植体培养能够利用光学显微镜和扫描电子显微镜研究隐孢子虫病的发病机制,并研究有无辣木叶提取物处理的组织中细胞因子水平的变化。该模型有助于理解肠道分泌和炎症反应的调节,可用于筛选潜在的抗隐孢子虫候选化合物。