Baicalin suppresses expression of Chlamydia protease-like activity factor in Hep-2 cells infected by Chlamydia trachomatis

In this study, we tested the ability of Baicalin to block Chlamydia trachomatis infection and found that the Baicalin blocked infection of Hep-2 cells. Then, we looked into the expression of RFX5 and CPAF gene in Chlamydia-infected cells. We found that RFX5 and CPAF were up-regulated and down-regulated respectively by Baicalin. Since CPAF is responsible for degrading RFX5, we suggest that CPAF was a primary target of Baicalin and played an important role in regulating RFX5. Our findings demonstrate that Baicalin can inhibit C. trachomatis effectively and therefore, can be considered as potential agents for therapy of Chlamydia infectious diseases.     

抗沙眼衣原体主要外膜蛋白(MOMP)单克隆抗体的制备及初步应用

制备抗沙眼衣原体(CT)单克隆抗体,建立沙眼衣原体的胶体金免疫层析快速检测方法。采用纯化沙眼衣原体主要外膜蛋白(MOMP)免疫Balb/c小鼠,选取高效价的免疫小鼠脾细胞与骨髓瘤细胞融合,ELISA检测筛选分泌抗MOMP单克隆抗体(McAb)的细胞株并进行相关鉴定。共获得分泌IgG1,IgG2b和IgM的3株单克隆抗体杂交瘤细胞株。纯化的单克隆抗体与鹦鹉热衣原体、肺炎衣原体、肺炎支原体均无交叉反应。通过位点分析选择配对良好的两株McAb,制备检测CT的双抗体夹心法免疫层析试纸条。制备的胶体金试纸条对MOMP的最低检出值可达50 ng·mL-1。     

Seroprevalence of chlamydial infection in dairy cattle in Guangzhou, southern China

Chlamydia spp. are obligate intracellular gram-negative bacteria that cause a wide range of significant diseases in humans and animals worldwide, resulting in significant economic losses. Chlamydial infection in cattle has been reported in many countries including China. However, there has been no survey of chlamydial infection of dairy cattle in Guangzhou, southern China. The objective of the present investigation was to examine the chlamydial seroprevalence in dairy cattle in Guangzhou, subtropical southern China by using an indirect hemagglutination assay (IHA). The overall seroprevalence of chlamydial infection in dairy cattle was 7.25% (29/400). Greater than or equal to eight-yr-old dairy cattle had the highest seroprevalence (10.34%), followed by those that were ≥ 6 years old or < 7 years old dairy cattle (10.20%), although there were no statistically significant differences among different groups (P > 0.05). Dairy cattle with 5 pregnancies had the highest seroprevalence (10.81%). These results indicate that chlamydial infection was present in dairy cattle in Guangzhou, subtropical southern China, and integrated strategies and measures should be executed to control and prevent chlamydial infection and disease outbreak in the study region.     

羊流产衣原体的分离鉴定及间接免疫荧光法检测

对收集的疑似衣原体感染的羊流产胎儿样本接种鸡胚卵黄囊进行分离培养,通过PCR-RFLP(限制性片段长度多态性分析)方法鉴定为流产衣原体。将收集到的流产衣原体阳性样本接种Hela229细胞,应用基于流产衣原体MOMP单克隆抗体的间接免疫荧光法,在Hela229细胞浆内可见呈绿色荧光的衣原体包涵体,进一步在病原学水平上确定发生在甘肃省肃南县羊流产的病原为流产衣原体。     

猪衣原体感染的血清学调查报告

通过对平顶山地区6个县(市)29个猪场的3000头种猪血清进行鹦鹉热衣原体的间接血凝(IHA)抗体的检测。结果显示,血清的阳性率为3.2%～8%,平均为4.8%,猪场的阳性率为65%,该病在本地区较为普遍。     

河北进境种用动物衣原体血清学检测

衣原体病是一种重要的人畜共患病,给养殖业带来巨大的经济损失,同时严重威胁从业人员的身体健康。本研究对河北辖区引进的2批种鸡、2批种鸭及2批种猪进行抽样,使用间接血凝试验（IHA）检测引进种用动物的衣原体抗体。结果表明：衣原体IHA抗体阳性率为15.7%。在种鸡、种鸭和种猪中均检测到衣原体IHA抗体,阳性率分别为18.1%、4.2%和20%。这说明在引进种用动物中存在着较高的衣原体抗体阳性率,需要加强对该病的检测。     

Infection of the Bovine Female Genital Tract with Chlamydia psittaci as a Possible Cause of Infertility

Contents: Chlamydia psittaci was isolated in embryonated chicken eggs via the yolk sac route from ten (16,7%) vaginal andlor endometrial mucosal scrapings of 60 slaughter cows. Simultaneous fecal shedding of chlamydiae was found in four animals. Chlamydial infections of the genital tract were frequent (p < 0,01) when there were endometrial inflammatory lesions together with the failure to detect other bacterial pathogens in the uterus.     

Seroprevalences for ovine enzootic abortion in Switzerland

Our aim was to assess the seroprevalence of Chlamydophila (Cd) abortus (Chlamydia psittaci serotype 1), denoted ovine enzootic abortion (OEA), in the Swiss sheep population. A competitive enzyme-linked immunosorbent assay (cELISA) was adapted for the investigation of pooled serum samples (pool approach) and receiver-operator characteristic (ROC) analysis was applied to define the cut-off of the pool approach. At a cut-off value of 30% inhibition, the flock-level pooled sensitivity and specificity were 92.9% and 97.6% when compared to classifying the flock based on individual-animal samples.

Subsequently, sera from 775 randomly selected flocks out of 11 cantons of Switzerland were investigated using the pool approach. The cantons included in the study represented 72% of the Swiss sheep flocks and 76% of Swiss sheep population. Antibodies against Cd. abortus were found in almost 19% (144) of the 775 examined sheep flocks. Test prevalences were adjusted for the imperfect test characteristics using the Rogan–Gladen estimator and Bayesian inference. Seroprevalence was highest (43%) in the canton Graubünden. In the remaining 10 cantons the seroprevalence ranged from 2 to 29%. The cELISA in combination with testing pooled sera and statistical methods for true prevalence estimation provided a good survey tool at lower costs and time when compared to other approaches.     

L929细胞无血清悬浮驯化机器在流产衣原体灭活疫苗中的应用研究

为了给规模化生产流产衣原体提供悬浮细胞培养方法,本研究通过逐步降血清悬浮驯化法使贴壁L929细胞逐步适应悬浮培养环境,最终获得了一株可无血清悬浮培养的L929细胞株;将流产衣原体接种于L929悬浮细胞上,研究不同的促感染试剂以提高收菌量,并选择出最佳的培养方法;制备流产衣原体灭活疫苗,以25 μL 、50 μL、100 μL的剂量免疫Balb/c小鼠,攻毒后检测其免疫保护效果。结果显示,当该L929悬浮细胞初始接种密度为5×105个/mL 时,培养72 h后细胞浓度可达到约7×106个/mL,细胞活力在95%以上;在细胞密度为6×106个/mL,接菌量为5.5×104 IFU/mL,加入脂质体2000的量为1.7 μL/mL、放线菌酮浓度为0.4 μg/mL时,菌量增殖倍数最大,48 h内约扩增了52倍。经流产衣原体悬浮细胞灭活疫苗免疫后,小鼠产生的抗体水平随免疫剂量的增加而提升;接种50 μL、100 μL疫苗组小鼠的脾淋巴细胞增殖水平、IFN-γ、IL-2的含量远高于接种25 μL疫苗和注射PBS组的小鼠;对免疫后的小鼠进行攻毒,接种疫苗组小鼠的脾脏、十二指肠、子宫中的流产衣原体基因组含量远低于未接种疫苗组小鼠。注射PBS组及接种25 μL疫苗组的小鼠子宫出现了不同程度的坏死及炎症,而其余两组小鼠的子宫无异常变化。结果表明,本试验成功驯化出１株L929悬浮培养细胞株,并且通过悬浮培养工艺制备的流产衣原体灭活疫苗免疫效果良好,接种50 μL、100 μL该灭活疫苗可以有效抑制流产衣原体的感染,该L929细胞全悬浮培养株的成功驯化为疫苗规模化生产提供了科学依据。     

Detection of Chlamydiaceae in ocular swabs from Australian pre‐export feedlot sheep

Infectious Ovine Keratoconjunctivitis (IOK) is a contagious ocular disease of sheep. A range of organisms have been observed as the aetiological agents of IOK. In this study, the presence of chlamydial pathogens (C. pecorum, C. abortus, C. psittaci) in conjunctival swabs was tested for. The swabs were collected from sheep with varying grades of IOK in an Australian pre‐export feedlot. The sheep had been rejected from a shipment because of the eye disease. The relative contribution of chlamydial pathogens to IOK and the rejection of animals was evaluated. In total, 149 conjunctival swabs were taken from rejected sheep (IOK Grades 1 to 6; n = 126) as well as those with healthy eyes (Grade 0; n = 23). Screening for chlamydial pathogens was done using species–specific qPCR assays. Chlamydial DNA was detected in 35.6% (53/149) of conjunctival samples. C. pecorum was the most predominant species with an overall prevalence of 28.9% (43/149). C. psittaci prevalence was 6.7% (10/149). Both organisms were detected in healthy as well as IOK‐affected eyes. All swabs tested negative for C. abortus. The results from this study demonstrate that Chlamydia spp can be readily detected in sheep presenting with IOK. The zoonotic C. abortus was not detected in any of the samples in this study, providing further evidence to the suggestion that this pathogen remains absent from Australia. Although the exact contribution of Chlamydia spp in the IOK pathogenesis is unclear, such studies are anticipated to be of benefit to Australian domestic and live export production systems.