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用牛型和羊型副结核分枝杆菌亚种对幼年马鹿( Cervus elaphus )进行实验性感染 。

Experimental infections in young red deer (Cervus elaphus) with a bovine and an ovine strain of Mycobacterium avium subsp paratuberculosis.

作者信息

Mackintosh C G, Labes R E, Clark R G, de Lisle G W, Griffin J F T

机构信息

AgResearch Invermay, PO Box 50034, Mosgiel, New Zealand.

出版信息

N Z Vet J. 2007 Feb;55(1):23-9. doi: 10.1080/00480169.2007.36731.

Abstract

AIMS

To compare the virulence of a 'bovine' and an 'ovine' strain of Mycobacterium avium subsp paratuberculosis (M. ptb) in red deer (Cervus elaphus) after experimental inoculation orally, and to examine the relationship between the dose of the bovine strain given and immunological, clinical and histopathological outcomes in young red deer.

METHODS

Newly-weaned 4-month-old male red deer (n=81) were randomly assigned to one of five groups. Three groups (n=16) received high (10(9) colony forming units (cfu); HB), medium (10(7) cfu; MB) or low (10(3) cfu; LB) oral doses of a bovine strain of M. ptb, one group (n=16) received medium (10(7) cfu; MO) doses of an ovine strain of M. ptb, and a Control group (n=17) was not dosed. The HB and Control groups were grazed together, the MB and LB groups were grazed together, and the MO group was grazed alone, in separate small paddocks on a quarantined area of the farm for 45 weeks. Liveweight, clinical signs and immunoglobulin G1 (IgG1) antibody levels were monitored for up to 45 weeks. Deer affected with Johne's disease were euthanised when they showed obvious clinical signs. Unaffected deer were slaughtered at the end of the trial (Week 45), and all deer were necropsied. Faeces and tissue samples were cultured for M. ptb, and fixed tissues were examined for histopathology.

RESULTS

Between 21 and 38 weeks post-challenge (pc), 5/16 animals in the HB group developed early signs of Johne's disease and were euthanised. The remaining deer in the five groups were all apparently healthy and reached good liveweights (approximately 100 kg average), and were euthanised and examined 45 weeks pc. Three deer (two HB and one MB) had small caseous lesions in their jejunal lymph nodes (JJLNs) and one HB animal had a small caseous lesion in a retropharyngeal lymph node. The remaining animals had no grossly-visible lesions. Mycobacterium avium subsp paratuberculosis was cultured from samples from 100% of the HB and MB animals, 50% of the LB group, 69% of the MO group and all Control animals. Thus all Control deer were infected by natural transmission from the HB group but none developed signs of clinical disease. Examination of histological sections of jejunum, ileocaecal valve (ICV) and associated lymph nodes showed a gradation of severity of lesions that was positively correlated (p<0.001) with dose of the bovine strain administered; mean lesion severity scores were 4.8, 2.9 and 0.9 for HB, MB and LB groups, and 2.2 and 0.9 for the Control and MO groups, respectively. IgG1 antibody levels at the time of euthanasia were also correlated with lesion severity scores at slaughter (p<0.001).

CONCLUSIONS

The ovine strain of M. ptb used in this study was less virulent for red deer than the bovine strain. The correlation between dose of the bovine strain and the severity of lesions suggests that clinical Johne's disease in yearling red deer likely results from high oral challenge with a bovine strain whilst they are young. The minimum oral infective dose may be close to 10(3) cfu for this bovine strain.

摘要

目的

比较经口实验接种后,“牛型”和“羊型”副结核分枝杆菌(M. ptb)菌株对马鹿(Cervus elaphus)的毒力,并研究给幼龄马鹿接种牛型菌株的剂量与免疫、临床及组织病理学结果之间的关系。

方法

将刚断奶的4月龄雄性马鹿(n = 81)随机分为五组。三组(n = 16)分别经口给予高剂量(10⁹ 菌落形成单位(cfu);HB)、中剂量(10⁷ cfu;MB)或低剂量(10³ cfu;LB)的牛型M. ptb菌株,一组(n = 16)经口给予中剂量(10⁷ cfu;MO)的羊型M. ptb菌株,对照组(n = 17)不接种。HB组和对照组一起放牧,MB组和LB组一起放牧,MO组单独放牧,在农场隔离区的单独小围栏中饲养45周。监测体重、临床症状和免疫球蛋白G1(IgG1)抗体水平长达45周。患有副结核病的马鹿出现明显临床症状时实施安乐死。未受影响的马鹿在试验结束时(第45周)屠宰,所有马鹿均进行剖检。对粪便和组织样本进行M. ptb培养,对固定组织进行组织病理学检查。

结果

攻毒后(pc)21至38周,HB组16只动物中有5只出现副结核病早期症状并实施安乐死。其余五组马鹿均明显健康,体重良好(平均约100 kg),并在攻毒后45周实施安乐死并进行检查。三只马鹿(两只HB组和一只MB组)空肠淋巴结(JJLNs)有小的干酪样病变,一只HB组动物咽后淋巴结有小的干酪样病变。其余动物无肉眼可见病变。100%的HB组和MB组动物样本、50%的LB组动物样本、69%的MO组动物样本以及所有对照组动物样本均培养出副结核分枝杆菌。因此,所有对照组马鹿均因与HB组自然接触而感染,但均未出现临床疾病症状。对空肠、回盲瓣(ICV)及相关淋巴结的组织学切片检查显示,病变严重程度呈梯度变化,与所接种牛型菌株的剂量呈正相关(p < 0.001);HB组、MB组和LB组的平均病变严重程度评分分别为4.8、2.9和0.9,对照组和MO组分别为2.2和0.9。安乐死时的IgG1抗体水平也与屠宰时的病变严重程度评分相关(p < 0.001)。

结论

本研究中使用的羊型M. ptb菌株对马鹿的毒力低于牛型菌株。牛型菌株剂量与病变严重程度之间的相关性表明,一岁龄马鹿的临床副结核病可能是由于幼年时经口高剂量接触牛型菌株所致。该牛型菌株的最低经口感染剂量可能接近10³ cfu。

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