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[临床铜绿假单胞菌菌株的抗念珠菌活性及对念珠菌生物膜形成的体外抑制作用]

[Anti-candidal activity of clinical Pseudomonas aeruginosa strains and in vitro inhibition of Candida biofilm formation].

作者信息

Keçeli Özcan Sema, Dündar Devrim, Sönmez Tamer Gülden

机构信息

Kocaeli University Faculty of Medicine, Department of Medical Microbiology, Kocaeli, Turkey.

出版信息

Mikrobiyol Bul. 2012 Jan;46(1):39-46.

Abstract

It has been shown recently that signal-mediated interactions between the opportunistic pathogens Pseudomonas aeruginosa and Candida albicans affect virulence features in both organisms. It has been emphasized that the anti-candidal activity of P.aeruginosa is especially due to its pyocyanin pigment. The structure and the properties of monospecies biofilms and their role in infectious disease have been extensively studied. However, there are only few studies on interactions between P.aeruginosa ve Candida spp. İn mixed biofilms. Therefore, in this study, the investigation of anti-candidal activity of P.aeruginosa against Candida spp. And the effect on C.albicans biofilm were aimed. A total of 58 P.aeruginosa strains isolated from several clinical specimens and one of each C.albicans ATCC 90028, C.parapsilosis ATCC 22019, C.glabrata ATCC 90030 and a C.tropicalis clinical isolate were included in the study. The anti-candidal activity of P.aeruginosa strains were investigated on Sabouraud dextrose agar using Kerr's method. The absence of the growth of Candida strains were evaluated as total inhibition. The effect of two P.aeruginosa strains showing total inhibition and one isolate without inhibition effect on C.albicans biofilm was investigated by microplate method. For the evaluation of monospecies or mixed species biofilms of Pseudomonas and C.albicans, the number of colony forming units (cfu/ml) after 90 minutes (adhesion phase), 24 hours and 48 hours (biofilm phase) incubation were determined. Then, cfu numbers of monospecies and mixed species of Pseudomonas and C.albicans were compared. Total inhibition rates of Pseudomonas strains against C.albicans, C.tropicalis, C.glabrata and C.parapsilosis were determined as 63.8%, 75.9%, 65.5% and 72.4%, respectively. A significant reduction was determined in cfu/ml numbers of mixed species of Pseudomonas and C.albicans compared to cfu numbers in monospecies biofilm after 90 minutes, 24 hours and 48 hours (p< 0.05). In addition to the inhibitory effect of P.aeruginosa on Candida growth, in a dual species environment both organisms mutually suppressed each others' biofilm development quantitatively. These findings indicated that molecular basis of signal-mediated interaction between Pseudomonas and Candida spp. Should be clarified for better understanding of the pathogenesis of mixed bacterial-fungal infections.

摘要

最近的研究表明,机会致病菌铜绿假单胞菌和白色念珠菌之间的信号介导相互作用会影响这两种生物体的毒力特征。有人强调,铜绿假单胞菌的抗念珠菌活性尤其归因于其绿脓菌素色素。单一物种生物膜的结构、特性及其在传染病中的作用已得到广泛研究。然而,关于铜绿假单胞菌与念珠菌属在混合生物膜中的相互作用的研究却很少。因此,在本研究中,旨在研究铜绿假单胞菌对念珠菌属的抗念珠菌活性以及对白色念珠菌生物膜的影响。本研究共纳入了从多个临床标本中分离出的58株铜绿假单胞菌菌株,以及白色念珠菌ATCC 90028、近平滑念珠菌ATCC 22019、光滑念珠菌ATCC 90030各1株和1株热带念珠菌临床分离株。采用克尔氏法在沙氏葡萄糖琼脂上研究铜绿假单胞菌菌株的抗念珠菌活性。念珠菌菌株生长的缺失被评估为完全抑制。通过微孔板法研究了两株表现出完全抑制作用的铜绿假单胞菌菌株和一株对白色念珠菌生物膜无抑制作用的分离株的影响。为了评估铜绿假单胞菌和白色念珠菌的单一物种或混合物种生物膜,测定了孵育90分钟(黏附阶段)、24小时和48小时(生物膜阶段)后的菌落形成单位数量(cfu/ml)。然后,比较了铜绿假单胞菌和白色念珠菌单一物种和混合物种的cfu数量。铜绿假单胞菌菌株对白色念珠菌、热带念珠菌、光滑念珠菌和近平滑念珠菌的总抑制率分别确定为63.8%、75.9%、65.5%和72.4%。与单一物种生物膜中的cfu数量相比,在90分钟、24小时和48小时后,铜绿假单胞菌和白色念珠菌混合物种的cfu/ml数量显著降低(p<0.05)。除了铜绿假单胞菌对念珠菌生长的抑制作用外,在双物种环境中,两种生物体在数量上相互抑制彼此的生物膜形成。这些发现表明,为了更好地理解混合细菌 - 真菌感染的发病机制,应阐明铜绿假单胞菌与念珠菌属之间信号介导相互作用的分子基础。

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