Konold Timm, Thorne Leigh, Simmons Hugh A, Hawkins Steve A C, Simmons Marion M, González Lorenzo
Animal Sciences Unit, Animal and Plant Health Agency Weybridge, New Haw, Addlestone, Surrey, KT15 3NB, UK.
Virology Department, Animal and Plant Health Agency Weybridge, New Haw, Addlestone, Surrey, KT15 3NB, UK.
BMC Vet Res. 2016 Sep 17;12:208. doi: 10.1186/s12917-016-0807-4.
Previous studies confirmed that classical scrapie can be transmitted via milk in sheep. The current study aimed to investigate whether scrapie can also be transmitted via goat milk using in vivo (new-born lambs fed milk from scrapie-affected goats due to the unavailability of goat kids from guaranteed scrapie-free herds) and in vitro methods (serial protein misfolding cyclic amplification [sPMCA] on milk samples).
In an initial pilot study, new-born lambs of two different prion protein gene (PRNP) genotypes (six VRQ/VRQ and five ARQ/ARQ) were orally challenged with 5 g brain homogenate from two scrapie-affected goats to determine susceptibility of sheep to goat scrapie. All sheep challenged with goat scrapie brain became infected based on the immunohistochemical detection of disease-associated PrP (PrP(sc)) in lymphoid tissue, with an ARQ/ARQ sheep being the first to succumb. Subsequent feeding of milk to eight pairs of new-born ARQ/ARQ lambs, with each pair receiving milk from a different scrapie-affected goat, resulted in scrapie in the six pairs that received the largest volume of milk (38-87 litres per lamb), whereas two pairs fed 8-9 litres per lamb, and an environmental control group raised on sheep milk from healthy ewes, did not show evidence of infection when culled at up to 1882 days of age. Infection in those 12 milk recipients occurred regardless of the clinical status, PrP(sc) distribution, caprine arthritis-encephalitis virus infection status and PRNP polymorphisms at codon 142 (II or IM) of the donor goats, but survival time was influenced by PRNP polymorphisms at codon 141. Serial PMCA applied to a total of 32 milk samples (four each from the eight donor goats collected throughout lactation) detected PrP(sc) in one sample each from two goats.
The scrapie agent was present in the milk from infected goats and was able to transmit to susceptible species even at early preclinical stage of infection, when PrP(sc) was undetectable in the brain of the donor goats. Serial PMCA as a PrP(sc) detection method to assess the risk of scrapie transmission via milk in goats proved inefficient compared to the bioassay.
先前的研究证实,经典羊瘙痒病可通过绵羊的乳汁传播。本研究旨在利用体内方法(由于无法获得来自无羊瘙痒病保证羊群的新生山羊,用受羊瘙痒病影响的山羊的乳汁喂养新生羔羊)和体外方法(对乳汁样本进行连续蛋白质错误折叠循环扩增[sPMCA]),研究羊瘙痒病是否也能通过山羊奶传播。
在一项初步的试点研究中,对两种不同朊病毒蛋白基因(PRNP)基因型的新生羔羊(6只VRQ/VRQ和5只ARQ/ARQ)口服接种来自两只受羊瘙痒病影响的山羊的5克脑匀浆,以确定绵羊对山羊羊瘙痒病的易感性。根据淋巴组织中与疾病相关的PrP(PrP(sc))的免疫组织化学检测,所有用山羊羊瘙痒病脑匀浆攻击的绵羊均被感染,其中一只ARQ/ARQ绵羊是第一只死亡的。随后,给8对新生ARQ/ARQ羔羊喂食乳汁,每对羔羊接受来自不同受羊瘙痒病影响的山羊的乳汁,结果显示,接受乳汁量最大的6对羔羊(每只羔羊38 - 87升)感染了羊瘙痒病,而另外两对每只羔羊喂食8 - 9升乳汁,以及一个用健康母羊的羊奶饲养的环境对照组,在1882日龄时宰杀时未显示感染迹象。这12只接受乳汁的羔羊感染与否与供体山羊的临床状态、PrP(sc)分布、山羊关节炎 - 脑炎病毒感染状态以及密码子142(II或IM)处的PRNP多态性无关,但存活时间受密码子141处的PRNP多态性影响。对总共32份乳汁样本(来自8只供体山羊在整个泌乳期各采集4份)应用连续PMCA,在来自两只山羊的各一份样本中检测到了PrP(sc)。
感染山羊的乳汁中存在羊瘙痒病病原体,即使在供体山羊脑内检测不到PrP(sc)的感染早期临床前期阶段,该病原体也能够传播给易感物种。与生物测定法相比,连续PMCA作为一种评估山羊乳汁传播羊瘙痒病风险的PrP(sc)检测方法被证明效率低下。