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Our research group has recently shown that Borrelia burgdorferi , the Lyme disease bacterium, is capable of forming biofilms in Borrelia-infected human skin lesions called Borrelia lymphocytoma (BL). Biofilm structures often contain multiple organisms in a symbiotic relationship, with the goal of providing shelter from environmental stressors such as antimicrobial agents. Because multiple co-infections are common in Lyme disease, the main questions of this study were whether BL tissues contained other pathogenic species and/or whether there is any co-existence with Borrelia biofilms. Recent reports suggested Chlamydia -like organisms in ticks and Borrelia -infected human skin tissues; therefore, Chlamydia -specific polymerase chain reaction (PCR) analyses were performed in Borrelia -positive BL tissues. Analyses of the sequence of the positive PCR bands revealed that Chlamydia spp. DNAs are indeed present in these tissues, and their sequences have the best identity match to Chlamydophila pneumoniae and Chlamydia trachomatis. Fluorescent immunohistochemical and in situ hybridization methods demonstrated the presence of Chlamydia antigen and DNA in 84% of Borrelia biofilms. Confocal microscopy revealed that Chlamydia locates in the center of Borrelia biofilms, and together, they form a well-organized mixed pathogenic structure. In summary, our study is the first to show Borrelia-Chlamydia mixed biofilms in infected human skin tissues, which raises the questions of whether these human pathogens have developed a symbiotic relationship for their mutual survival.
Our research group has recently shown that Borrelia burgdorferi , the Lyme disease bacterium, is capable of forming biofilms in Borrelia-infected human skin lesions called Borrelia lymphocytoma (BL). Biofilm structures often contain multiple organisms in a symbiotic relationship, with the goal of providing shelter from environmental stressors such as antimicrobial agents. Because multiple co-infections are common in Lyme disease, the main questions of this study were whether BL tissues contained other pathogenic species and/or whether there is any co-existence with Borrelia biofilms. Recent reports suggested Chlamydia -like organisms in ticks and Borrelia -infected human skin tissues; therefore, Chlamydia -specific polymerase chain reaction (PCR) analyses were performed in Borrelia -positive BL tissues. Analyses of the sequence of the positive PCR bands revealed that Chlamydia spp. DNAs are indeed present in these tissues, and their sequences have the best identity match to Chlamydophila pneumoniae and Chlamydia trachomatis. Fluorescent immunohistochemical and in situ hybridization methods demonstrated the presence of Chlamydia antigen and DNA in 84% of Borrelia biofilms. Confocal microscopy revealed that Chlamydia locates in the center of Borrelia biofilms, and together, they form a well-organized mixed pathogenic structure. In summary, our study is the first to show Borrelia-Chlamydia mixed biofilms in infected human skin tissues, which raises the questions of whether these human pathogens have developed a symbiotic relationship for their mutual survival.
Community-acquired pneumonia (CAP) is a common disease responsible for significant morbidity and mortality. However, the definite etiology of CAP often remains unresolved, suggesting that unknown agents of pneumonia remain to be identified. The recently discovered members of the order Chlamydiales, Chlamydia-related bacteria (CRB), are considered as possible emerging agents of CAP. Parachlamydia acanthamoebae is the most studied candidate. It survives and replicates inside free-living amoeba, which it might potentially use as a vehicle to infect animals and humans. A Mycoplasma pneumoniae outbreak was observed in Kymenlaakso region in Southeastern Finland during August 2017–January 2018. We determined the occurrence of Chlamydiales bacteria and their natural host, free-living amoeba in respiratory specimens collected during this outbreak with molecular methods. Altogether, 22/278 (7.9%) of the samples contained Chlamydiales DNA. By sequence analysis, majority of the CRBs detected were members of the Parachlamydiaceae family. Amoebal DNA was not detected within the sample material. Our study further proposes that Parachlamydiaceae could be a potential agent causing atypical CAP in children and adolescents.
Introduction: Waddlia chondrophila and Parachlamydia acanthamoebae are well-known and best-studied representatives of Сhlamydia-related bacteria carrying a potential zoonotic threat. These bacteria are associated with miscarriage, ectopic pregnancy, diseases of the respiratory system in both humans and animals. Despite the importance of these Сhlamydia-like organisms for human medicine along with veterinary medicine, studies on their prevalence in Ukraine were not conducted due to the lack of available tests. The aim of our work was to create relatively cheap and easy method for detection Waddlia chondrophila and Parachlamydia acanthamoebae. Materials and methods: GenBank database was used to find nucleotide sequences of the 16S rRNA gene of bacteria Chlamydiales’ order. Alignment was performed using the MEGA7 software, in order to detect the presence of polymorphic hybridization sites specifically attributed to Waddlia chondrophila and Parachlamydia acanthamoebae. Primer- BLAST software was used to design oligonucleotide primers, to evaluate the critical parameters of the primer, in particular, the melting temperature, difference between melting temperatures for the primer pairs, the GC content, the self-complementarity, etc. Results and conclusions: The amplification of control DNA of Parachlamydia acanthamoebae and Waddlia chrondophila in single PCR using the corresponding primers and subsequent gel electrophoresis of PCR products determined the size of the amplified DNA fragments 88 b.p. and 123 b.p, respectively; the fragments were in line with the expected sizes. The analytical specificity test was performed by amplifying the control DNA of 15 species of the order Chlamydiales.
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