Chlamydophila psittaci

披肩衣原体
  • 文章类型: Journal Article
    鹦鹉衣原体-鸟类中的人畜共患病原体-可能会传播给人类,导致严重的呼吸道疾病。在家禽养殖场工作或居住在家禽养殖场附近的个人极易感染鹦鹉。在这项研究中,我们通过收集来自不同家禽品种的粪便样本和来自家禽农场和动物园工人的咽拭子样本以及血清样本,评估了鹦鹉在中国三个城市的家禽和人类中的流行和基因型。通过靶向C.psittaciompA的实时荧光定量PCR(qPCR)筛选这些样品。对阳性样品进行PCR扩增和ompA测序。根据ompA序列的系统发育分析,对样品中检测到的菌株进行基因分型。总的来说,3.13%(40/1278)的家禽粪便样本在qPCR检测中呈阳性,而在qPCR和间接荧光抗体检测中,3.82%(6/157)的咽拭子样本和42.59%(46/108)的工人血清样本呈阳性,分别。在32个家禽样品和6个人类样品中检测到的菌株被基因分型为A型,表明工人感染了产于农场禽类的鹦鹉。此外,8只孔雀菌株的基因型为CPX0308,在中国首次鉴定。阐明鹦鹉在动物和家禽相关工人中的分布可能为降低种群中鹦鹉感染的风险提供有价值的见解。
    Chlamydia psittaci-a zoonotic pathogen in birds-may be transmitted to humans, causing severe respiratory disease. Individuals working in or living near poultry farms are highly susceptible to C. psittaci infection. In this study, we assessed the prevalence and genotypes of C. psittaci in poultries and humans in three cities of China by collecting fecal samples from different poultry species and throat swab samples and serum samples from workers in poultry farms and zoos. These samples were screened by real-time fluorescence quantitative PCR (qPCR) targeting C. psittaci ompA. The positive samples were subjected to PCR amplification and sequencing of ompA. The strains detected in the samples were genotyped on the basis of the phylogenetic analysis of ompA sequences. In total, 3.13% (40/1278) poultry fecal samples were positive in the qPCR assay, whereas 3.82% (6/157) of throat swab samples and 42.59% (46/108) of serum samples from the workers were positive in the qPCR and indirect fluorescent antibody assays, respectively. The strains detected in the 32 poultry samples and 6 human samples were genotyped as type A, indicating that the workers were infected with C. psittaci that originated in poultry birds in farms. Additionally, eight peacocks showed strains with the genotype CPX0308, which was identified in China for the first time. Elucidating the distribution of C. psittaci in animals and poultry-related workers may provide valuable insights for reducing the risk of C. psittaci infection within a population.
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  • 文章类型: Case Reports
    这项研究介绍了一名74岁的男性患者因20天的咳嗽史而入院的临床资料,胸闷,和呼吸困难。一被录取,病人出现发烧,心动过速,和呼吸急促.临床检查显示肺部感染的证据,脓毒症,和多器官功能障碍,除了异常的血气分析和升高的C反应蛋白(CRP)水平。病原体检测证实鹦鹉衣原体(C.Psittaci),感染。在整个治疗过程中,患者并发真菌和病毒感染,需要一种涉及抗生素和抗真菌联合治疗的综合方法。尽管遇到了治疗相关的并发症,患者在积极管理下表现出临床改善.该病例强调了识别衣原体感染后免疫抑制的重要性,强调早期诊断的关键作用,干预,以及提高患者预后的标准化治疗方案。
    This study presents the clinical profile of a 74-year-old male patient admitted to the hospital due to a 20-day history of coughing, chest tightness, and dyspnea. Upon admission, the patient presented with fever, tachycardia, and tachypnea. Clinical examination revealed evidence of lung infection, sepsis, and multi-organ dysfunction, alongside abnormal blood gas analysis and elevated C-reactive protein (CRP) levels. Pathogen testing confirmed Chlamydia psittaci (C. psittaci), infection. Throughout the treatment course, the patient developed concurrent fungal and viral infections, necessitating a comprehensive approach involving combined antibiotic and antifungal therapy. Despite encountering treatment-related complications, the patient demonstrated clinical improvement with aggressive management. This case underscores the importance of recognizing immune suppression subsequent to Chlamydia infection, emphasizing the critical role of early diagnosis, intervention, and standardized treatment protocols in enhancing patient prognosis.
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  • 文章类型: Journal Article
    细胞内细菌从宿主细胞和细胞组织中的排出是感染周期中的关键过程。这个过程对于细菌在宿主内部传播是必不可少的,并且可以影响感染的结果。对于专性细胞内革兰氏阴性人畜共患病细菌鹦鹉衣原体,对细菌从感染的上皮中排出的机制知之甚少。这里,我们描述和表征含衣原体的球体(CCS),衣原体利用的一种新型且主要的非裂解性出口。CCS是球形的,低相差结构被具有特定屏障功能的磷脂酰丝氨酸暴露膜包围。它们含有感染性后代和形态受损的细胞器。CCS的形成是一个连续的过程,从含有DEVD四肽的底物的蛋白水解裂解开始,可以在衣原体内含物内部检测到,随后感染细胞的细胞内钙浓度增加。随后,质膜开始起泡,包涵膜不稳定,并且含有DEVD的底物的蛋白水解裂解在整个感染细胞内迅速增加。最后,感染,泡状细胞分离并离开单层,从而形成CCS。该事件序列对于衣原体CCS形成是独特的,并且与先前描述的衣原体外出途径根本不同。因此,CCS的形成代表了一个主要的,先前未表征的胞内病原体的出口途径,通常可能与衣原体生物学有关,并可能影响体内感染结果。重要宿主细胞外出对于细胞内病原体在生物体内传播和宿主到宿主的传播是必不可少的。这里,我们将含衣原体球体(CCS)的形成描述为细胞内病原体鹦鹉衣原体和沙眼衣原体的新型且主要的非裂解性出口途径。CCS的形成与挤压形成有根本的不同,先前描述的沙眼衣原体的非裂解出口途径。CCS的形成是一个独特的连续过程,包括蛋白水解活性,随后细胞内钙浓度增加,包涵膜不稳定,质膜起泡,和整个磷脂酰丝氨酸暴露的前宿主细胞的最终分离。因此,CCS的形成代表了细胞内病原体的重要且先前未表征的出口途径,这可能与衣原体生物学有关。包括寄主向性,保护宿主细胞防御机制,或细菌致病性。
    The egress of intracellular bacteria from host cells and cellular tissues is a critical process during the infection cycle. This process is essential for bacteria to spread inside the host and can influence the outcome of an infection. For the obligate intracellular Gram-negative zoonotic bacterium Chlamydia psittaci, little is known about the mechanisms resulting in bacterial egress from the infected epithelium. Here, we describe and characterize Chlamydia-containing spheres (CCSs), a novel and predominant type of non-lytic egress utilized by Chlamydia spp. CCSs are spherical, low-phase contrast structures surrounded by a phosphatidylserine-exposing membrane with specific barrier functions. They contain infectious progeny and morphologically impaired cellular organelles. CCS formation is a sequential process starting with the proteolytic cleavage of a DEVD tetrapeptide-containing substrate that can be detected inside the chlamydial inclusions, followed by an increase in the intracellular calcium concentration of the infected cell. Subsequently, blebbing of the plasma membrane begins, the inclusion membrane destabilizes, and the proteolytic cleavage of a DEVD-containing substrate increases rapidly within the whole infected cell. Finally, infected, blebbing cells detach and leave the monolayer, thereby forming CCS. This sequence of events is unique for chlamydial CCS formation and fundamentally different from previously described Chlamydia egress pathways. Thus, CCS formation represents a major, previously uncharacterized egress pathway for intracellular pathogens that could be linked to Chlamydia biology in general and might influence the infection outcome in vivo.IMPORTANCEHost cell egress is essential for intracellular pathogens to spread within an organism and for host-to-host transmission. Here, we characterize Chlamydia-containing sphere (CCS) formation as a novel and predominant non-lytic egress pathway of the intracellular pathogens Chlamydia psittaci and Chlamydia trachomatis. CCS formation is fundamentally different from extrusion formation, the previously described non-lytic egress pathway of C. trachomatis. CCS formation is a unique sequential process, including proteolytic activity, followed by an increase in intracellular calcium concentration, inclusion membrane destabilization, plasma membrane blebbing, and the final detachment of a whole phosphatidylserine-exposing former host cell. Thus, CCS formation represents an important and previously uncharacterized egress pathway for intracellular pathogens that could possibly be linked to Chlamydia biology, including host tropism, protection from host cell defense mechanisms, or bacterial pathogenicity.
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  • 文章类型: Journal Article
    该研究的目的是描述鹦鹉病肺炎,并评估C反应蛋白/白蛋白比值对鹦鹉病肺炎严重程度的预测价值。
    使用宏基因组测序诊断的鹦鹉病肺炎病例数据来自上海的三家医院,中国从10月2019年10月2022年。测定血清C-反应蛋白和白蛋白水平,并计算C-反应蛋白与白蛋白的比值(CAR)。斯皮尔曼相关分析,有序逻辑回归分析,并进行受试者工作特征曲线分析,考察3项指标对病情严重程度的相关性和预测能力。
    共纳入27例鹦鹉病肺炎患者,平均年龄62岁,70.4%为男性。44.4%的患者有明确的禽类或禽类接触史。主要症状为发热(100%)。在呼吸重症监护病房(RICU)接受治疗的患者发生喘息的可能性更高(88.9%对33.3%,P=0.013)和胸闷(88.9%vs.33.3%,P=0.013)高于普通病房(非RICU)。RICU中胸腔积液患者的比例明显高于非RICU(88.9%vs.38.9%,P=0.019)。RICU组的CAR明显高于非RICU组(9.41vs.4.05,P=0.017)。这一结果伴随着更高的插管和呼吸机支持(33.3%vs.0.0%,P=0.029),RICU的PCT和CRP水平高于非RICU,白蛋白和PaCO2水平较低。Logistic回归分析显示,CAR(OR1.49;95%CI1.07~2.06,P=0.017)是住院时间延长(>14天)的危险因素。
    研究发现,血清CAR水平升高与严重鹦鹉病肺炎的风险增加相关。因此,它可以作为一个简单和有用的诊断工具,为临床医生迅速和准确地确定的严重程度的鹦鹉病肺炎,最终帮助他们制定最佳的治疗方案。
    UNASSIGNED: The aim of the study was to describe psittacosis pneumonia and to assess the predictive value of the C-reactive protein/albumin ratio in psittacosis pneumonia for severity.
    UNASSIGNED: Data on psittacosis pneumonia cases diagnosed using metagenomic sequencing were collected from three hospitals in Shanghai, China from Oct. 2019 to Oct. 2022. Serum levels of C-reactive protein and albumin were measured and the C-reactive protein to albumin ratio (CAR) was calculated. Spearman\'s correlation analysis, ordered logistic regression analysis, and receiver operating characteristic curve analysis were conducted to examine the correlation and predictive ability of the three indicators on the severity of the disease.
    UNASSIGNED: A total of 27 patients with psittacosis pneumonia were enrolled, with an average age of 62 years and 70.4% being male. 44.4% of patients had a clear history of contact with poultry or birds. The predominant symptom was fever (100%). Patients treated in the respiratory intensive care unit (RICU) had a higher likelihood of experiencing wheezing (88.9% versus 33.3%, P=0.013) and chest tightness (88.9% vs. 33.3%, P=0.013) than those in the general ward (Non-RICU). The proportion of patients with pleural effusion was significantly higher in the RICU compared to the Non-RICU (88.9% vs. 38.9%, P=0.019). The RICU group had a significantly higher CAR than the Non-RICU group (9.41 vs. 4.05, P=0.017). This result was accompanied by higher intubation and ventilator support (33.3% vs. 0.0%, P=0.029), higher PCT and CRP levels and lower albumin and PaCO2 levels in the RICU than in the Non-RICU. Logistic regression analysis indicated that CAR (OR 1.49; 95% CI 1.07-2.06, P=0.017) was risk factor for prolonged hospitalization (> 14 days).
    UNASSIGNED: Elevated serum CAR levels were found to be associated with a greater risk of severe psittacosis pneumonia. Consequently, it may serve as an uncomplicated and useful diagnostic tool for clinicians to promptly and precisely ascertain the severity of psittacosis pneumonia, ultimately aiding them in devising the most optimal therapeutic plan.
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  • 文章类型: Journal Article
    背景:我们调查了2021-2022年长沙市活禽批发市场家禽中鹦鹉衣原体的存在和环境,并进行了系统发育分析,以了解其在该市场的分布。
    方法:总共,使用实时聚合酶链反应分析了483个样品,使用高通量测序分析了17个C。惊人的相似性,和系统发育分析。
    结果:在483个家禽和环境样本中,有22个对鹦鹉梭菌呈阳性(总阳性率:4.55%),在12个月内的阳性率没有差异。在11个采样点检测到鹦鹉衣原体(总阳性率:27.5%),包括鸡肉,鸭子,和鸽子/鸡/鸭/鹅店,鸽子店的阳性率最高(46.67%)。污水中阳性率最高(12.5%),家禽粪便(7.43%),笼式拭子(6.59%),禽咽/阴囊拭子(3.33%),和空气(2.29%)样品。ompA序列在两个鹦鹉的菌株中进行了鉴定,使用系统发育分析确定具有基因型B。因此,在监测过程中,在长沙市家禽批发市场的家禽和环境样品中检测到B型C.psittaci基因型。
    结论:为了应对潜在的人畜共患威胁,C.应加强活禽市场的鹦鹉鱼监测计划。
    BACKGROUND: We investigated the presence of Chlamydia psittaci in poultry and the environment in live poultry wholesale markets in Changsha during 2021-2022 and conducted a phylogenetic analysis to understand its distribution in this market.
    METHODS: In total, 483 samples were analyzed using real-time polymerase chain reaction and 17 C. psittaci-positive samples using high-throughput sequencing, BLAST similarity, and phylogenetic analysis.
    RESULTS: Twenty-two out of 483 poultry and environmental samples were positive for C. psittaci (overall positivity rate: 4.55%) with no difference in positivity rates over 12 months. Chlamydia psittaci was detected at 11 sampling points (overall positivity rate: 27.5%), including chicken, duck, and pigeon/chicken/duck/goose shops, with pigeon shops having the highest positivity rate (46.67%). The highest positivity rates were found in sewage (12.5%), poultry fecal (7.43%), cage swab (6.59%), avian pharyngeal/cloacal swab (3.33%), and air (2.29%) samples. The ompA sequences were identified in two strains of C. psittaci, which were determined to bear genotype B using phylogenetic analysis. Thus, during monitoring, C. psittaci genotype B was detected in the poultry and environmental samples from the poultry wholesale market in Changsha.
    CONCLUSIONS: To address the potential zoonotic threat, C. psittaci monitoring programs in live poultry markets should be enhanced.
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  • 文章类型: Journal Article
    鹦鹉衣原体肺炎(CPP)是由鹦鹉衣原体感染引起的肺部疾病。这可能导致严重的急性呼吸窘迫综合征和全身症状。本研究探讨了活性氧(ROS)对CPP中Th17/Treg平衡影响的具体机制。采用ELISA和流式细胞术检测健康个体和CPP患者外周血中ROS水平和Th17/Treg分化比,分别。使用Pearson相关分析评估ROS水平与Th17/Treg之间的关联。在H2O2处理和NLRP3抑制后,测量CD4+T细胞中的ROS水平和Thl7/Treg比率。使用免疫印迹法观察H2O2处理和NLRP3抑制对NLRP3/IL-1β/caspase-1通路的影响。与健康组相比,CPP组外周血中ROS水平升高,Th17分化率升高,Treg分化率降低。ROS水平与Th17细胞比例呈正相关,与Treg细胞比例呈负相关。H2O2处理后CD4+T细胞中ROS水平和NLRP3/IL-1β/caspase-1表达上调。此外,Th17分化增加,Treg分化减少。相反,NLRP3/IL-1β/caspase-1通路的抑制逆转了H2O2治疗的作用,ROS水平没有显著变化。ROS调节CPP中的Th17/Treg平衡,可能通过NLRP3/IL-1β/caspase-1途径。本研究为CPP免疫治疗的发展提供了新的视角。
    Chlamydia psittaci pneumonia (CPP) is a lung disease caused by the infection with the Chla-mydia psittaci bacterium, which can lead to severe acute respiratory distress syndrome and systemic symptoms. This study explored the specific mechanisms underlying the impact of reactive oxygen species (ROS) on the Th17/Treg balance in CPP. The levels of ROS and the differentiation ratio of Th17/Treg in the peripheral blood of healthy individuals and CPP patients were measured using ELISA and flow cytometry, respectively. The association between the ROS levels and Th17/Treg was assessed using Pearson correlation analysis. The ROS levels and the Th17/Treg ratio were measured in CD4+ T cells following H2O2 treatment and NLRP3 inhibition. The effects of H2O2 treatment and NLRP3 inhibition on the NLRP3/IL-1β/caspase-1 pathway were observed using immunoblotting. Compared to the healthy group, the CPP group exhibited increased levels of ROS in the peripheral blood, an elevated ratio of Th17 differentiation, and a decreased ratio of Treg differentiation. ROS levels were positively correlated with the Th17 cell proportion but negatively correlated with the Treg cell proportion. The ROS levels and NLRP3/IL-1β/caspase-1 expression were up-regulated in CD4+ T cells after H2O2 treatment. Furthermore, there was an increase in Th17 differentiation and a decrease in Treg differentiation. Conversely, the NLRP3/IL-1β/caspase-1 pathway inhibition reversed the effects of H2O2 treatment, with no significant change in the ROS levels. ROS regulates the Th17/Treg balance in CPP, possibly through the NLRP3/IL-1β/caspase-1 pathway. This study provides a new perspective on the development of immunotherapy for CPP.
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  • 文章类型: Case Reports
    背景技术在COVID-19大流行期间,快速诊断对于有效治疗重症肺炎至关重要。披肩衣原体,一种非典型的社区获得性病原体,通常表现出非特异性的临床体征,需要严格的微生物培养条件,复杂的识别。宏基因组下一代测序(mNGS),涉及到从临床样本中对DNA或RNA进行鸟枪测序,是临床环境中的一项关键技术。尽管mNGS技术能够识别核酸,它不应该直接等同于病原微生物的存在。尽管如此,它有望成为未来检测严重传染病中非典型病原体的主要方法。病例报告我们介绍了一例严重的社区获得性鹦鹉衣原体肺炎,突出了COVID-19的持续突变和频繁传播。患者的严重肺部感染迅速进展,导致多器官衰竭,需要体外膜氧合(ECMO)支持。尽管最初的常规实验室测试尚无定论,通过mNGS证实了鹦鹉衣原体肺炎的诊断。给予抗生素治疗和多器官功能支持,导致患者的成功康复和出院。结论在COVID-19大流行期间,诊断由非典型病原体引起的重症肺炎面临重大挑战。在没有有效感染控制的情况下启动ECMO支持会带来相当大的风险,如导管相关感染和抗菌治疗失败的风险增加。在这个案例中,mNGS被证明有助于筛选严重传染病中的非典型病原体。多器官功能支持在可逆条件下的应用为临床医生提供了鉴定和治疗病原体的时间。为在疫情爆发期间诊断和治疗由非常规病原体引起的重症肺炎提供了新的方法。
    BACKGROUND Rapid diagnosis is critical for effective treatment of severe pneumonia during the COVID-19 pandemic. Chlamydia psittaci, an atypical community-acquired pathogen, typically exhibits nonspecific clinical signs and requires stringent conditions for microbiological culture, complicating its identification. Metagenomic next-generation sequencing (mNGS), which involves shotgun sequencing of DNA or RNA from clinical samples, is a key technology in clinical settings. Although mNGS technology identifies nucleic acids, it should not be directly equated with the presence of pathogenic microorganisms. Nonetheless, it shows promise as a principal method for detecting atypical pathogens in severe infectious diseases in the future. CASE REPORT We present a case of severe community-acquired Chlamydia psittaci pneumonia, highlighting the ongoing mutations and frequent spread of COVID-19. The patient\'s severe pulmonary infection rapidly advanced, resulting in multiple organ failure, necessitating extracorporeal membrane oxygenation (ECMO) support. Despite initial inconclusive routine laboratory tests, diagnosis of Chlamydia psittaci pneumonia was confirmed through mNGS. Antibiotic treatment and multi-organ functional support were administered, leading to the patient\'s successful recovery and hospital discharge. CONCLUSIONS Diagnosing severe pneumonia caused by atypical pathogens amid the COVID-19 pandemic presents significant challenges. Initiating ECMO support without effective infection control poses considerable risks, such as increasing risk of catheter-related infections and antimicrobial treatment failure. In the case presented, mNGS proved instrumental in screening for atypical pathogens in critical infectious diseases. Application of multi-organ function support in reversible conditions affords clinicians time for pathogen identification and treatment, offering novel approaches for diagnosing and treating severe pneumonia induced by unconventional pathogens during epidemic outbreaks.
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  • 文章类型: Case Reports
    在一名没有病史的年轻人中,诊断出与鹦鹉衣原体相关的社区获得性肺炎与急性心肌炎相关。和对鸟类的专业博览会。支气管肺泡灌洗特异性聚合酶链反应阳性证实了诊断。患者接受螺旋霉素治疗2周,同时抗炎治疗心肌炎3个月。快速观察到临床和生物学改善,随后心电图和胸部CT扫描恢复正常。在超过两年的随访中没有报告复发。
    Chlamydia psittaci ‒ related community-acquired pneumonia associated to acute myocarditis was diagnosed in a young man with no medical history, and a professional exposition to birds. The diagnosis was confirmed with positive specific polymerase chain reaction in bronchoalveolar lavage. The patient was treated with spiramycin for two weeks with anti-inflammatory treatment for myocarditis for three months. Clinical and biological improvement was rapidly observed followed by normalization of electrocardiogram and chest CT scan. No relapse was reported for over a two-year follow-up.
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  • 文章类型: Journal Article
    不同的综合征涉及人类鹦鹉病(流感样综合征,非典型肺炎直至泪腺淋巴瘤)。诊断方法包括血清学,文化,和PCR。暴露工人中的鹦鹉衣原体(Cp)阳性检测率仍然未知。我们的研究旨在评估与布宜诺斯艾利斯自然保护区中的载体鸟类接触的工人的阳性测试率,阿根廷。次要目的是分析与这些结果相关的风险因素以及表明鹦鹉病的体征的发生。鼻咽拭子和血清对是从与确认的携带者鸟类互动的员工那里收集的。具有可检测的Cp和/或抗衣原体属DNA的那些。抗体基线滴度≥160mUI/ml,或者至少四倍,被认为是积极的。与鸟类一起或在鸟类附近进行的活动,个人防护设备的使用,和以前的慢性条件进行评估。在随访期间评估了可能的Cp相关病理。总共对63名暴露工人(71.4%的男性)进行了评估,中位年龄为35.7岁(IQR26-39),检测到28.6%的阳性。慢性呼吸系统疾病是与阳性测试相关的独特因素(OR5.2[1.5-18.5]p<.05)。令人惊讶的是,大约三分之一的工人结果是阳性,所有人都对医疗做出了反应,没有人发展与典型的鹦鹉病相关的急性非典型肺炎综合征。作为人畜共患疾病预防措施的一部分,强烈建议对接触携带者或潜在携带者鸟类的动物工作者进行积极的早期诊断和适当的治疗。
    Different syndromes are involved in human psittacosis (flu-like syndrome, atypical pneumonia up to lacrimal gland lymphoma). Diagnostic methods include serology, culture, and PCR. The rate of Chlamydia psittaci (Cp) positive tests among exposed workers is still unknown. Our study aimed to assess the rate of positive tests among workers who have contact with carrier birds in natural reserves from Buenos Aires, Argentina. Secondary aims were to analyze risk factors linked to these outcomes and the occurrence of signs that suggest psittacosis. Nasopharyngeal swabs and serum pairs were collected from employees who had interacted with confirmed carrier birds. Those with detectable DNA of Cp and/or anti-Chlamydia spp. antibody baseline titer ≥ 160 mUI/ml, or at least quadruplicating, were considered positive. Activities performed with or near birds, personal protective equipment use, and previous chronic conditions were assessed. Possible Cp-related pathologies were evaluated during follow-up. A total of 63 exposed workers (71.4% men) with a median age of 35.7 years (IQR 26-39) were evaluated to detect 28.6% positives. Respiratory chronic conditions were the unique factor associated with positive tests (OR 5.2 [1.5-18.5] p < .05). Surprisingly, about a third of the workers resulted positive and all responded to medical treatment, none developing an acute atypical pneumonia syndrome associated with classical presentation of psittacosis. Active testing for early diagnosis and proper treatment in zoological workers exposed to carrier or potentially carrier birds is strongly suggested as part of zoonotic diseases preventive measures.
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  • 文章类型: Journal Article
    鹦鹉衣原体是一种禽类细菌病原体,可通过人畜共患传播引起人类非典型肺炎。它是一种革兰氏阴性细胞内细菌,在活真核细胞的细胞质中的膜结合内含物内部增殖。在不破坏这种细胞的情况下对这种细胞进行研究提出了重大挑战。我们在这项工作中证明了组合的多工具方法分析此类复杂样品的实用性。应用原子力显微镜在细菌进入时获得受感染细胞表面的高分辨率图像。原子力显微镜扫描显示衣原体感染细胞的细胞膜形态变化,例如细胞膜粗糙度的变化和微囊泡的存在。使用4Pi拉曼显微镜对完整细胞内胞内细菌的分子组成进行成像和探测。获得了有关C.psittaci产生的内含物结构的信息,发现其分子指纹与细胞内脂滴相似,但蛋白质和不饱和脂质较少。所提出的方法证明了各种基于显微镜的方法的互补性,并且可能对细胞内细菌的表征有用。
    Chlamydia psittaci is an avian bacterial pathogen that can cause atypical pneumonia in humans via zoonotic transmission. It is a Gram-negative intracellular bacterium that proliferates inside membrane bound inclusions in the cytoplasm of living eukaryotic cells. The study of such cells with C. psittaci inside without destroying them poses a significant challenge. We demonstrated in this work the utility of a combined multitool approach to analyze such complex samples. Atomic force microscopy was applied to obtain high-resolution images of the surface of infected cells upon entrance of bacteria. Atomic force microscopy scans revealed the morphological changes of the cell membrane of Chlamydia infected cells such as changes in roughness of cell membrane and the presence of micro vesicles. 4Pi Raman microscopy was used to image and probe the molecular composition of intracellular bacteria inside intact cells. Information about the structure of the inclusion produced by C. psittaci was obtained and it was found to have a similar molecular fingerprint as that of an intracellular lipid droplet but with less proteins and unsaturated lipids. The presented approach demonstrates complementarity of various microscopy-based approaches and might be useful for characterization of intracellular bacteria.
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