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猪的实验性弓形虫感染:体液免疫反应、特异性IgG亲和力评估以及母猪垂直传播再现的挑战

Experimental Toxoplasma gondii infections in pigs: Humoral immune response, estimation of specific IgG avidity and the challenges of reproducing vertical transmission in sows.

作者信息

Basso Walter, Grimm Felix, Ruetten Maja, Djokic Vitomir, Blaga Radu, Sidler Xaver, Deplazes Peter

机构信息

Institute of Parasitology, Vetsuisse-Faculty, University of Zurich, Winterthurerstrasse 266a, CH-8057 Zurich, Switzerland; Department of Farm Animals, Division of Swine Medicine, Vetsuisse-Faculty, University of Zurich, Winterthurerstrasse 260, CH-8057 Zurich, Switzerland.

Institute of Parasitology, Vetsuisse-Faculty, University of Zurich, Winterthurerstrasse 266a, CH-8057 Zurich, Switzerland.

出版信息

Vet Parasitol. 2017 Mar 15;236:76-85. doi: 10.1016/j.vetpar.2017.01.026. Epub 2017 Feb 3.

Abstract

Ten pregnant sows were experimentally inoculated per os with T. gondii in order to investigate vertical and galactogenic transmission of the parasite and the evolution and maturation of the specific IgG humoral response in the sows and piglets. Five seronegative sows received 10T. gondii (CZ isolate clone H3) sporulated oocysts during late-pregnancy (Exp. 1), three sows received 10 oocysts during mid-pregnancy (Exp. 2) and three sows from Exp. 1 (and two seronegative sows) were re-inoculated with 10 oocysts during a further pregnancy (late-pregnancy) (Exp. 3). Besides, six 4.5 week-old piglets inoculated per os with 5×10 oocysts were also included in the serological investigations. All animals seroconverted (PrioCHECK Toxoplasma Ab porcine ELISA, Prionics, Switzerland) by 2-3 weeks post inoculation (wpi) and remained seropositive for at least 38 weeks or until euthanasia. Four chronically infected sows from Exp. 1 and 2 were serologically monitored during a further pregnancy and no reactivation, but a decrease of the antibody levels was observed at farrowing (Exp. 4). In all experiments, the specific IgG-avidity was initially low, increased during the course of infection and after re-inoculations. An avidity index (AI) ≥40% could be used to rule out recent infections (<8 weeks) in most (15 of 16) animals. In some piglets (18.6% of 70) delivered by inoculated sows (Exp. 1 and 2), maternal antibodies were still detectable at 2 months (but not by 3 months) of age, with constant high avidity values, comparable to those of the dams at farrowing. In all experiments, the sows remained asymptomatic and delivered non-infected offspring at term. A total of 208 normal and 5 stillborn piglets delivered by the inoculated sows (Exp. 1-4) tested serologically negative before colostrum uptake. Placentas (n=88) from all sows and tissues (brain, liver, lung, heart, and masseter muscle) from 56 delivered piglets were analysed histopathologically and by real-time PCR for T. gondii with negative results. Colostrum and milk samples from all sows were negative by real-time PCR for T. gondii DNA. In addition, no seroconversion was observed in 16 piglets from seronegative dams that were transferred to infected dams one day after birth to detect a possible infection through colostrum or milk during the suckling period. Although vertical transmission of T. gondii was demonstrated in naturally infected pigs, many factors involved in the outcome of vertical transmission and congenital toxoplasmosis in pigs are still unknown.

摘要

为了研究弓形虫的垂直传播和经乳汁传播、母猪和仔猪体内特异性IgG体液免疫反应的演变及成熟情况,对10头怀孕母猪进行了经口接种弓形虫的实验。5头血清学阴性的母猪在妊娠后期接受了10个弓形虫(CZ分离株克隆H3)孢子化卵囊(实验1),3头母猪在妊娠中期接受了10个卵囊(实验2),实验1中的3头母猪(以及2头血清学阴性的母猪)在再次怀孕(妊娠后期)时再次接种了10个卵囊(实验3)。此外,6头4.5周龄经口接种5×10个卵囊的仔猪也纳入了血清学研究。所有动物在接种后2 - 3周(wpi)血清转化(采用瑞士Prionics公司的PrioCHECK猪弓形虫抗体ELISA检测),并至少38周保持血清阳性或直至安乐死。对实验1和2中4头慢性感染的母猪在再次怀孕期间进行血清学监测,未观察到再激活,但在分娩时抗体水平下降(实验4)。在所有实验中,特异性IgG亲和力最初较低,在感染过程中及再次接种后升高。在大多数(16头中的15头)动物中,亲和力指数(AI)≥40%可用于排除近期感染(<8周)。在接种母猪(实验1和2)所产的一些仔猪(70头中的18.6%)中,在2月龄时仍可检测到母源抗体(但3月龄时未检测到),其亲和力值持续较高,与分娩时母猪的亲和力值相当。在所有实验中母猪均无症状,足月分娩出未感染的后代。接种母猪(实验1 - 4)所产的20%正常仔猪和5头死产仔猪在摄取初乳前血清学检测为阴性。对所有母猪的胎盘(n = 88)以及56头分娩仔猪的组织(脑、肝、肺、心和咬肌)进行了组织病理学分析和弓形虫实时PCR检测,结果均为阴性。所有母猪的初乳和乳汁样本弓形虫DNA实时PCR检测均为阴性。此外,将出生一天后从血清学阴性母猪转至感染母猪处的16头仔猪在哺乳期间未观察到血清转化,以检测通过初乳或乳汁可能发生的感染。虽然在自然感染的猪中已证实存在弓形虫的垂直传播,但猪垂直传播和先天性弓形虫病结果所涉及的许多因素仍不清楚。

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