Bakırcı Serkan, Bilgiç Hüseyin Bilgin, Köse Onur, Aksulu Ayça, Hacılarlıoğlu Selin, Karagenç Tülin, Çavuş İbrahim, Özbilgin Ahmet
Adnan Menderes Üniversitesi Veteriner Tıp Fakültesi, Parazitoloji Anabilim Dalı, Aydın, Türkiye.
Turkiye Parazitol Derg. 2015 Sep;39(3):212-7. doi: 10.5152/tpd.2015.4300.
This study aimed to observation the possible visceralization tendency and dissemination of L. major amastigotes in gerbils (Meriones unguiculatus) using a classic smear technique, inoculated into enriched Novy-MacNeal-Nicolle (NNN) culture and polymerase chain reaction (PCR) assay for diagnosis of infection.
In this study, L. major isolated from a man who 18 years old, living in Bitlis province of Turkey. This strain also was utilized to infect gerbils. A total of 1 × 10(8)/mL promastigotes were inoculated to 10 gerbils. Necropsy was performed on infected gerbils for monitoring the visceralization tendency of the parasites. Tissue samples were prepared from each animal and stained by Giemsa and inoculated into NNN culture. However, a real-time PCR assay was performed to confirm the infection the clinical material.
Examination of Giemsa-stained tissue smears showed that infected animals with L.major were positive for Leishmania amastigotes in all tissues at the first month post infection and Leishmania promastigotes were cultured at 26°C in culture flasks containing NNN. Melting curve analyses of ribozomal internal transcribed spacer 1 (ITS-1) PCR showed the peak concordant with L. major.
As a result, the present study confirmed by both Giemsa-stained smears and PCR, visceralization and dissemination of L. major amastigotes, the principal cause of zoonotic cutaneous leishmaniasis in gerbils.
本研究旨在通过经典涂片技术观察大沙鼠（Meriones unguiculatus）体内硕大利什曼原虫无鞭毛体的可能内脏化倾向及播散情况，将其接种于改良诺维 - 麦克尼尔 - 尼科尔（NNN）培养基并采用聚合酶链反应（PCR）检测法诊断感染情况。
在本研究中，硕大利什曼原虫分离自一名18岁居住在土耳其比特利斯省的男子。该菌株也被用于感染大沙鼠。将总共1×10⁸/mL前鞭毛体接种到10只大沙鼠体内。对感染的大沙鼠进行尸检以监测寄生虫的内脏化倾向。从每只动物身上制备组织样本，用吉姆萨染色并接种到NNN培养基中。然而，采用实时PCR检测法来确认临床材料中的感染情况。
吉姆萨染色组织涂片检查显示，感染硕大利什曼原虫的动物在感染后第一个月所有组织中的利什曼原虫无鞭毛体均呈阳性，并且利什曼原虫前鞭毛体在含有NNN的培养瓶中于26°C培养。核糖体内部转录间隔区1（ITS - 1）PCR的熔解曲线分析显示峰值与硕大利什曼原虫一致。
因此，本研究通过吉姆萨染色涂片和PCR均证实了硕大利什曼原虫无鞭毛体在大沙鼠体内的内脏化及播散情况，而硕大利什曼原虫是大沙鼠人兽共患皮肤利什曼病的主要病因。
