Tatman Otkun Müşerref, Akçalı Alper, Karadenizli Aynur, Ozbey Nilgün, Gazel Deniz, Sener Alper, Güçlü Oğuz, Tanrıöver Arzu, Otkun Metin
Çanakkale Onsekiz Mart University Faculty of Medicine, Department of Medical Microbiology, Çanakkale, Turkey.
Mikrobiyol Bul. 2011 Jan;45(1):48-57.
Tularemia is a disease caused by Francisella tularensis and widely seen at northern hemisphere of the world. In Turkey, oropharyngeal infections caused by a less virulent serotype F.tularensis subsp. Holarctica are more prevalent. The aim of this study was to present the results of an epidemiological research performed after the detection of tularemia cases from Biga county of Canakkale province, Turkey, in December 2009. Following the report of two tularemia suspected cases from two villages (Baliklicesme and Sinekci) of Biga, an epidemiological investigation was undertaken to inspect the situation in this area. Water samples, clinical samples as throat swabs, wound swabs and serum samples were collected. Samples were cultured on heart agar supplemented with sheep blood, cysteine and antibiotics. Cultures were incubated at 37°C in 5% CO(2) and followed for 10 days. Suspected colonies were identified by slide agglutination test using F.tularensis antisera. F.tularensis antibodies were investigated by standard tube agglutination method. Positive results obtained with agglutination test were also checked for a probable crossreaction with Brucella antibodies by Rose-Bengal test. Water and wound samples were investigated using real-time polymerase chain reaction (RT Taqman PCR; Quantica, Techne Inc, UK) with probe and primers specific for ISFtu2 gene. All of the cultures yielded negative results, however eight of 16 water samples, one lymph node aspirate and one throat sample were found positive in F.tularensis TaqMan RT-PCR test. In tube agglutination test positive antibody titers between 1:20-1:1280 were detected in 36 of 115 serum samples. Two cases with antibody titers of 1:1280 and accompanying acute clinical findings, were diagnosed as tularemia and treated accordingly. Lymphatic drainage fluid samples obtained from one of these patients yielded positive result in PCR, however clinical sample could not be obtained from the other patient. The only epidemiological linkage between these acute cases (n= 2) and the other seropositive subjects (n= 34) was the use of local water supply system. It was learned that water obtained through reverse osmosis system had been used as drinking water at Baliklicesme village. Pre- and post-reverse osmosis system water samples from Baliklicesme village and samples from water supply of Sinekci village revealed positive results for F.tularensis by PCR. Since the only epidemiological relation between these two villages was using local water supply, tularemia cases encountered in this area were attributed to a waterborne epidemic and an automatic chlorination system was set up at each water reservoir in these villages. The establishment of these preventive measures curbed the growth of the epidemic. The cases presenting with throat sore, fever, lymphadenopathy (more than 2 cm), non-responsive to beta-lactam antibiotics, should be further investigated for tularemia. This work emphasizes that systematic setup and control of water disinfection systems are crucial to prevent tularemia outbreaks. Community and related authorities should be educated about the importance of water sanitation and chlorination.
兔热病是一种由土拉弗朗西斯菌引起的疾病,在世界北半球广泛存在。在土耳其,由毒性较低的土拉弗朗西斯菌亚种全北区血清型引起的口咽感染更为普遍。本研究的目的是呈现2009年12月在土耳其恰纳卡莱省比加县检测到兔热病病例后进行的一项流行病学研究结果。在比加的两个村庄(巴利克利切梅和锡内克奇)报告了两例疑似兔热病病例后,开展了一项流行病学调查以检查该地区的情况。采集了水样、临床样本如咽拭子、伤口拭子和血清样本。样本在补充有羊血、半胱氨酸和抗生素的心脏琼脂上培养。培养物在37°C、5%二氧化碳环境中孵育10天。使用土拉弗朗西斯菌抗血清通过玻片凝集试验鉴定疑似菌落。通过标准试管凝集法检测土拉弗朗西斯菌抗体。凝集试验获得的阳性结果还通过虎红试验检查是否可能与布鲁氏菌抗体发生交叉反应。使用针对ISFtu2基因的特异性探针和引物,通过实时聚合酶链反应(RT Taqman PCR;Quantica,Techne Inc,英国)对水和伤口样本进行检测。所有培养均产生阴性结果,然而在16份水样中的8份、1份淋巴结穿刺液和1份咽拭子样本中,土拉弗朗西斯菌TaqMan RT-PCR检测呈阳性。在115份血清样本中的36份中,试管凝集试验检测到阳性抗体滴度在1:20 - 1:1280之间。两例抗体滴度为1:1280并伴有急性临床症状的病例被诊断为兔热病并相应治疗。其中一名患者的淋巴引流液样本在PCR检测中呈阳性,然而另一名患者无法获取临床样本。这些急性病例(n = 2)与其他血清学阳性受试者(n = 34)之间唯一的流行病学关联是使用当地供水系统。据了解,巴利克利切梅村已将通过反渗透系统获得 的水用作饮用水。巴利克利切梅村反渗透系统前后的水样以及锡内克奇村供水样本经PCR检测,土拉弗朗西斯菌呈阳性。由于这两个村庄之间唯一的流行病学关联是使用当地供水,该地区出现的兔热病病例归因于水源性流行,并且在这些村庄的每个水库都设置了自动氯化系统。这些预防措施的建立遏制了疫情的发展。对于出现咽痛、发热、淋巴结肿大(超过2厘米)且对β-内酰胺抗生素无反应的病例,应进一步调查是否为兔热病。这项工作强调,系统设置和控制水消毒系统对于预防兔热病暴发至关重要。应向社区和相关当局宣传水卫生和氯化的重要性。