羊副结核性腹泻病病原体分离及药敏试验

为了诊断与治疗阿拉善盟阿左旗发生的羊腹泻病,试验对从阿拉善盟阿左旗采集的病料进行病原体分离培养、抗酸染色、革兰染色、ELISA检测,并对分离到的菌株进行药敏试验。结果表明:从肠系膜淋巴结分离到抗酸染色阳性、革兰染色阳性的短粗杆状副结核杆菌,2份血清中均有与副结核杆菌抗原结合的相应抗体的存在;分离菌对环丙沙星和卡那霉素敏感,对头孢呋肟和阿奇霉素中度敏感,对其他药物均产生不同程度的耐药性。     

Mycobacteria in aquarium fish: results of a 3‐year survey indicate caution required in handling pet‐shop fish

Fish are commonly infected with non‐tuberculous mycobacteria (NTM), which should be regarded as potential pathogens when handling aquarium fish and equipment. This study examined 107 aquarium fish from pet shops. Cultivation of the fish samples using different selective media was conducted for identification of NTM. Isolates were identified using the GenoType Mycobacterium common mycobacteria and additional species assays, sequencing of the 16S rRNA and rpoB genes, and real‐time PCR assay for identification of Mycobacterium (M.) marinum. Among the investigated fish, 79.4% (85/107) were positive for mycobacteria, with 8.2% (7 of 85) having two mycobacterial species present. Among the positive fish, the common pathogens M. marinum, Mycobacterium fortuitum (M. fortuitum group) and Mycobacterium chelonae were identified in approx. 90% of fish and other NTM species in 10%, including Mycobacterium peregrinum/septicum, Mycobacterium gordonae, Mycobacterium arupense, Mycobacterium kansasii, Mycobacterium ulcerans and Mycobacterium setense. The well‐known human pathogen M. marinum was present in 10.6% of the positive fish (9 of 85). The species of mycobacteria identified in the study are not only recognized as aquarium fish pathogens, but can also cause pathology in humans. Microbiological and clinical communities should therefore be sensitized to the role of NTM in infections associated with exposure to aquarium fish.     

Antibiotic treatment of zebrafish mycobacteriosis: tolerance and efficacy of treatments with tigecycline and clarithromycin

Zebrafish (Danio rerio) are a popular model organism used in a growing number of research fields. Maintaining healthy, disease‐free laboratory fish is important for the integrity of many of these studies. Mycobacteriosis is a chronic bacterial infection caused by several Mycobacterium spp. and is the second most common disease found in laboratory zebrafish. Current mycobacteriosis control measures recommend the removal of infected fish and in severe outbreaks, depopulation. These measures can be effective, but less disruptive measures should be assessed for controlling mycobacteriosis, particularly when valuable and rare lines of fish are affected. Here, the in vivo efficacy of two drug candidates, tigecycline (1 μg g^?1) and clarithromycin (4 μg g^?1), was tested in adult zebrafish experimentally infected with Mycobacterium chelonae. We assessed both short (14 day)‐ and long‐term (30 day) treatments and evaluated fecundity and pathological endpoints. Fecundity and histology results show that zebrafish tolerated antibiotics. Antibiotic treatments did not significantly impact the prevalence of acid‐fast granulomas; however, the severity of infections (acid‐fast granuloma intensity) was significantly decreased following treatments.     

Experimental exposure of zebrafish, Danio rerio (Hamilton), to Mycobacterium marinum and Mycobacterium peregrinum reveals the gastrointestinal tract as the primary route of infection: a potential model for environmental mycobacterial infection

The natural route by which fish become infected with mycobacteria is unknown. Danio rerio (Hamilton) were exposed by bath immersion and intubation to Mycobacterium marinum and Mycobacterium peregrinum isolates obtained from diseased zebrafish. Exposed fish were collected over the course of 8 weeks and examined for the presence of mycobacteriosis. Mycobacteria were consistently cultured from the intestines, and often from the livers and spleens of fish exposed by both methods. Mycobacteria were not observed in the gills. Histological analysis revealed that fish infected with M. marinum often developed granulomas accompanied by clinical signs of mycobacteriosis, while infection with M. peregrinum infrequently led to clinical signs of disease. Passage of the bacteria through environmental amoebae (Acanthamoeba castellani) was associated with increased growth of M. peregrinum over the course of 8 weeks, when compared to infection with the bacteria not passed through amoebae. The results provide evidence that zebrafish acquire mycobacteria primarily through the intestinal tract, resulting in mycobacterial dissemination.     

Availability of culture filtrate protein-10 (CFP-10) secreted from Mycobacterium pseudoshottsii for mycobacteriosis diagnosis in ginbuna crucian carp Carrasius auratus langsdorfii

Mycobacteriosis in cultured fish is a challenge for the aquaculture industry worldwide. Treatment by chemical administration is difficult and no effective vaccine has been developed. Therefore, detection and isolation by early diagnosis are important for prevention of the spread of the disease. In mammals, interferon gamma release assays have been established for detection of tuberculosis; these tests are based on the delayed-type hypersensitivity (DTH) response against culture filtrate protein-10 (CFP-10) and the 6-kDa early secreted antigen target (ESAT-6) of Mycobacterium tuberculosis. On the other hand, little is known about the fish immune response against the ESAT-6 and CFP-10 proteins of mycobacteria, although these responses should find application in the diagnosis of mycobacteriosis in fish. In the present study, we identified ESAT-6 and CFP-10 from Mycobacterium pseudoshottsii and cloned the corresponding genes. Intraperitoneal injection of the corresponding DNA plasmid constructs in ginbuna crucian carp yielded increased expression of the fish interferon-γ1-1-encoding gene (IFN-γ1-1). In contrast, IFN-γ1-1 expression accompanied by DTH response was observed only in the CFP-10-DNA plasmid-injected fish. Furthermore, fish that had been prophylactically injected with CFP-10-DNA plasmid exhibited increased survival of M. pseudoshottsii infection. Taken together, these results suggested that CFP-10 may facilitate diagnosis of mycobacteriosis.     

羊副结核病的致病特点及综合防控措施

羊副结核病是由副结核分枝杆菌引起的一种以顽固性腹泻、肠粘膜显著增厚为主要临床特征的慢性传染病,多呈现隐形感染,该病潜伏期长,传播迅速,呈全球性分布,对畜牧业的危害巨大。本文立足羊副结核病研究进展的实际情况,梳理总结了目前学术界对羊副结核病病原学、流行病学、致病机制、临床诊断和免疫综合防控等方面的研究讨论,为羊副结核病的诊断与防控提供了理论基础。在此基础上提出防控羊副结核病政府需制定有效政策,养殖场需加强饲养管理,形成科学有效的防控系统,为我国畜牧业的可持续健康发展提供了理论支撑。     

Anti-TB activity of Evodia elleryana bark extract

An ethyl acetate extract of bark from Evodia elleryana produced significant growth inhibition of Mycobacterium tuberculosis at concentrations only minimally inhibitory to human T cells. The crude extract yielded 95% inhibition of TB at 50 microg/ml. The crude extract yielded 29% growth inhibition of human T-cells in culture at that concentration.     

牛结核杆菌mpb-83基因的扩增和表达

采用PCR方法扩增牛结核杆菌mpb-83基因,并将其连接到T3克隆载体,然后克隆到原核表达载体PET-32a中,构建重组质粒PET-32a—mpb-83。以重组质粒转化大肠杆菌BL21,用NI柱纯化,最后纯化产物在SDS—PAGE中检测。结果得到约664bp的目的片段,且MPB-83可在大肠杆菌BL21中高效表达。     

Experimental Infection with Mycobacterium Avium,Serotype 2, in Pigs: 3. Oral Infection with Small Doses of M. Avium*

Pigs were inoculated orally with Mycobacterium avium in doses ranging from 15.6 × 10² to 15.6 × 10⁶ viable units daily for 15 days (Table 1). The animals were necropsied 31–32 days after the last inoculation.Pigs given doses of 15.6 × 10⁶ and 15.6 × 10⁵ viable units showed delayed hypersensitivity to avian tuberculin 24 days after the last inoculation (Table 2). The pig inoculated with 15.6 × 10⁶ viable units showed macro- and/or microscopic lesions of the intestines and the liver, and of the mandibular, mesenteric and hepatic lymph nodes. Cultures showed growth of M. avium from the same tissues and from the spleen and the left tracheobronchial lymph node. The pig inoculated with 15.6 × 10⁵ viable units showed a pronounced granulomatous infiltration in the tonsils and the mesenteric lymph nodes. Growth of M. avium was obtained from the tonsils, the intestinal mucosa (Peyer patch) and the mandibular and mesenteric lymph nodes. Viable unit counts were high in the tonsils and in the mesenteric lymph nodes (Tables 3 and 4).Lower doses gave rise to a minimal tissue reaction and/or very low viable unit counts, and are not considered to be capable of producing a progressive tuberculosis.