猪密螺旋体痢疾的诊断方法研究进展

猪痢疾是由猪痢疾密螺旋体引起的一种肠道传染病,其特征是大肠黏膜发生卡他性炎、出血性炎,继而引发纤维素性坏死性炎,呈现黏液性、出血性下痢,该病属二类传染病,给养猪业造成很大损失,临床上,已成为危害养猪业比较严重的传染病之一,从病原学诊断、流行病学诊断、临床诊断、血清学诊断、分子生物学诊断和病理学诊断等方面加以综述。     

The frequent detection of a treponeme in bovine digital dermatitis by immunocytochemistry and polymerase chain reaction

A study was carried out to determine whether spirochaetes are frequently associated with digital dermatitis in United Kingdom (UK) dairy cattle. Histopathological examination of lesions using a silver stain showed a large number of unidentified spirochaete-like organisms present in digital dermatitis hoof skin tissue in all examined biopsies. Immunocytochemical staining demonstrated that spirochaetes in skin lesions were identified by polyclonal antisera to Borrelia burgdorferi, Treponema denticola and Treponema vincentii (again all biopsies were positively stained), whereas monoclonal antibodies to B. burgdorferi and any Treponema pallidum did not stain any organisms in all biopsies. A PCR of 16S rRNA, previously shown to be specific for a new treponeme, was employed and produced positive results from 82.4% of digital dermatitis tissues. It is concluded that this spirochaete (or related spirochaetes), which is similar to human oral treponemes, is frequently associated with, and may be responsible for, pathological changes in digital dermatitis.     

A filter-assisted culture method for isolation of Treponema spp. from bovine digital dermatitis and their identification by MALDI-TOF MS

Digital dermatitis (DD) is a major infectious foot disease of cattle worldwide. Some DD stages are associated with lameness, and the disease has significant economic and animal welfare consequences. The pathogenesis of the disease is not yet fully understood, but Treponema spp. have been associated consistently with clinical cases. Isolation of these fastidious bacteria is difficult and cumbersome. We describe an improved method enabling the culturing of the 3 Treponema spp. (T. pedis, T. phagedenis, and T. medium) from bovine foot specimens derived from DD lesions, using a combination of membrane filtering and subsequent growth on selective agar media. The entire procedure from sampling to verification of individual Treponema spp. takes up to 24 d. In addition, we established a MALDI-TOF MS–based identification method to be applied for confirmation of the different Treponema spp. This scheme provides an unambiguous, simple, and straightforward identification procedure for DD-associated Treponema spp.     

猪痢疾密螺旋体的冻干保存试验

将密螺旋体菌株(包括猪痢疾密螺旋体和无害螺旋体)接种于华农1或3号血液琼脂平板上,置CO_2:H_2=20:80的环境中和37℃下培养5天。选择生长最好的琼脂平板,每个加入3ml PBS(0.01mol,pH7.2),洗下培养物,集中后加入等容积的S. H. 保护剂并使成均一的悬液分装安瓿瓶。用Edwards EF03型冻干机进行冷冻真空干燥,在真空状态下密封安瓿瓶,冻干菌种保存于4℃冰箱或-10～-15℃冰盒中。保存期:用新鲜保护剂者为8-13个月;用干燥保护剂者为二年。     

Physiological properties and classification of strains of Treponema sp. isolated from pigs in poland

Fifty-one treponemas were isolated from pigs. Twenty-three isolates with typical morphology and growth characteristic were beta hemolytic, enteropathogenic, produced indole and with exception of three strains did not ferment fructose. These strains were classified as typical T. hyodysenteriae and were usually isolated from pigs with symptoms of mucohemorrhagic diarrhoea. The seventeen other isolates were weakly beta hemolytic after 48 h incubation, enteropathogenic, 12 out of 17 produced indole, 10 out 17 fermented fructose. These strains were usually isolated from pigs with symptoms of gray-green diarrhoea and classified as T. hyodysenteriae 2 biotype or intermediate type. They may be compared with Treponema sp. isolated by Taylor et al. Eleven non enteropathogenic strains showed typical characteristic for T. innocens. Gas chromatography analysis of the fatty acids production from glucose, showed that all isolated treponemas produced acetate and butyrate. Typical T. hyodysenteriae produced additionally propionate. Strains of T. hyodysenteriae biotype 2 produced propionate or isobutyrate as well.     

福建省第一次发生猪痢疾的诊断及扑灭经过报告

本文首次报告了1985年10月福建省发生猪痢疾及其扑灭经过。根据临诊观察、病理变化和实验室检查结果做出诊断后,采取全群扑杀,彻底消毒、空关猪场四个月的严格防疫措施,消灭了这个猪痢疾的疫点。     

Transcellular penetration of Treponema phagedenis isolated from papillomatous digital dermatitis in polarized normal human epidermal keratinocytes in vitro

Papillomatous digital dermatitis (PDD) is a polymicrobial infection causing lameness in dairy cattle. Culture-independent analysis has shown that Treponema phagedenis is present consistently and predominantly in the lesions. However, the pathogenesis of PDD, especially the tissue penetration pathway, has not been examined. In the present study, we investigated whether T. phagedenis strains isolated from PDD produce proteolytic enzyme (s) for disruption of the epithelial cell barrier and have the ability to translocate in polarized normal human epidermal keratinocytes (NHEK) in vitro. Ten strains of T. phagedenis isolated from lesions did not show proteolytic activity on modified skim milk agar, although a human strain of T. denticola used as a control showed such activity. The integrity of tight junctions was monitored by measurement of transepithelial electrical resistance (TER). The TER values after inoculation of the T. phagedenis strains examined did not change during the experimental period; however, apical to basolateral translocation of T. phagedenis was confirmed after 24 hr by microscopy and Treponema-specific PCR. We further confirmed that translocation of T. phagedenis was accelerated by co-inoculation with live T. denticola, but not with heat-killed organisms. Furthermore, tight junction ZO-1 protein was not lost intensity after inoculation with T. phagedenis and the organism was observed in NHEK cells using a florescence microscope. These results suggest that T. phagedenis strains may translocate via a transcellular route in vitro and that the invasion is accelerated by other bacteria, such as T. denticola, producing proteolytic activity.