16S rRNA gene-based identification of cultured bacterial flora from host-seeking Ixodes ricinus, Dermacentor reticulatus and Haemaphysalis concinna ticks, vectors of vertebrate pathogens

2009 ◽  
Vol 54 (5) ◽  
pp. 419-428 ◽  
Author(s):  
I. Rudolf ◽  
J. Mendel ◽  
S. Šikutová ◽  
P. Švec ◽  
J. Masaříková ◽  
...  
2010 ◽  
Vol 139 (4) ◽  
pp. 524-529 ◽  
Author(s):  
E. TIJSSE-KLASEN ◽  
L. J. JAMESON ◽  
M. FONVILLE ◽  
S. LEACH ◽  
H. SPRONG ◽  
...  

SUMMARYA preliminary study was conducted to determine the presence of spotted fever rickettsiae in two species of British tick (Ixodes ricinusandDermacentor reticulatus). The 16S rRNA gene ofRickettsiaspp. was detected in 39/401 (9·7%) of ticks tested, including 22/338 (6·5%)I. ricinusand 17/63 (27%)D. reticulatus. Some positiveI. ricinussamples showed 100% homology withRickettsia helvetica(10/22), and most positiveD. reticulatusshowed 100% homology withR. raoultii(13/17). Five otherRickettsiaspp. were detected exhibiting 96–99% homology. Ticks positive for rickettsiae were collected from various hosts and from vegetation from eight counties across Great Britain. The distribution ofR. helveticain various engorged and unfed stages ofI. ricinussuggests thatR. helveticais widespread.R. raoultiiwas found in questing adultD. reticulatusin Wales and England. This is the first evidence of potentially pathogenic spotted fever rickettsiae in British ticks.


2018 ◽  
Author(s):  
John S Lambert ◽  
Michael John Cook ◽  
John Eoin Healy ◽  
Ross Murtagh ◽  
Gordana Avramovic ◽  
...  

Lyme borreliosis is a systemic infection caused by tick-borne pathogenic borreliae of the Borrelia burgdorferi sensu lato complex or of the more heterogeneous relapsing fever borrelia group. Clinical distinction of the infections due to different borrelia species is difficult. Accurate knowledge of the prevalence and the species of borreliae in the infected ticks in the endemic areas is valuable for formulating appropriate guidelines for proper management of this infectious disease. The purpose of this research was to design a readily implementable protocol to detect the divergent species of borreliae known to exist in Europe, using Irish samples of Ixodes ricinus ticks as the subject for study. Questing I. ricinus nymph samples were taken at six localities within Ireland. The crude DNA of each dried tick was extracted by hot NH4OH and used to initiate a same-nested PCR with a pair of borrelial genus-specific primers to amplify a highly conserved 357/358 bp segment of the 16S rRNA gene for detection and as the template for Sanger sequencing. To distinguish B. garinii from B. burgdorferi and to discriminate the various strains of B. garinii, a second 282 bp segment of the 16S rRNA gene was amplified for Sanger sequencing. A signature segment of the DNA sequence excised from the computer-generated electropherogram was submitted to the GenBank for BLAST alignment analysis. A 100% ID match with the unique reference sequence in the GenBank was required for the molecular diagnosis of the borrelial species or strain. We found the overall rate of borrelial infection in the Irish tick population to be 5%, with a range from 2% to 12% depending on the locations of tick collection. At least 3 species, namely B. garinii, B. valaisiana and B. miyamotoi, are infecting the ticks collected in Ireland. The isolates of B. garinii were confirmed to be strain BgVir, strain Bernie or strain T25. Since antigens for diagnostic serology tests may be species- or even strain-specific, expanded surveillance of the species and strains of the borreliae among human-biting ticks in Ireland is needed to ensure that the antigens used for the serology tests do contain the epitopes matching the antibodies elicited by the borrelial species and strains in the ticks cohabitating in the same environment.


2021 ◽  
Author(s):  
Shin Watanabe ◽  
Tadashi Shinoda ◽  
Masaaki Minagawa ◽  
Mari Toya ◽  
Daisuke Motooka ◽  
...  

Abstract Background: Scientists previously believed that bacteria could not grow in bile, which is rich in bacteriostatic compounds such as bile acids. Therefore, the biliary tract was considered to be a sterile environment. However, high-throughput (i.e., amplicon) sequencing analysis methods have recently been used to discover the bacterial flora in gallbladder bile in brain-dead patients with normal gallbladders. In a bacterial flora analysis targeting the 16S ribosomal gene, a specific flora was present in the bile of normal gallbladders. However, these results were not obtained from truly healthy individuals. Therefore, the aim of this study was to analyze the microbial flora of bile collected from pathologically normal gallbladders that were surgically removed from patients with hepatobiliary pancreatic diseases who had normal liver function.Results: All 12 bile samples obtained from the gallbladders had negative culture results, although a bacterial flora was detected in all samples via 16S ribosome gene analysis. The composition of the bacterial flora was very simple, and the Firmicutes, Proteobacteria, and Actinobacteria phyla were identified in all samples. Based on 16S rRNA gene profile analysis, the composition ratio accounted for more than 80% of the total number of reads. The Anaerobacillus, Delftia, Bacillus, Ralstonia, Ochrobactrum, Acidovorax, and Curvibacter genera were detected in all 12 samples. Based on 16S rRNA gene profile analysis, at the genus level, Anaerobacillus and Delftia accounted for 58.62%–87.63% of the identified bacteria in each sample. Conclusions: In this study, the bacterial flora in the gallbladder bile was not diverse. Contrary to previous reports, few bacteria belonging to the Bacteroidetes phylum were detected. The functional significance of the gallbladder bacterial flora requires further investigation.


PLoS ONE ◽  
2019 ◽  
Vol 14 (4) ◽  
pp. e0209881 ◽  
Author(s):  
John S. Lambert ◽  
Michael John Cook ◽  
John Eoin Healy ◽  
Ross Murtagh ◽  
Gordana Avramovic ◽  
...  

2012 ◽  
Vol 2012 ◽  
pp. 1-11 ◽  
Author(s):  
Naoto Sano ◽  
Yoshio Yamashita ◽  
Kazumasa Fukuda ◽  
Hatsumi Taniguchi ◽  
Masaaki Goto ◽  
...  

Intracystic fluid was aseptically collected from 11 patients with postoperative maxillary cyst (POMC), and DNA was extracted from the POMC fluid. Bacterial species were identified by sequencing after cloning of approximately 580 bp of the 16S rRNA gene. Identification of pathogenic bacteria was also performed by culture methods. The phylogenetic identity was determined by sequencing 517–596 bp in each of the 1139 16S rRNA gene clones. A total of 1114 clones were classified while the remaining 25 clones were unclassified. A total of 103 bacterial species belonging to 42 genera were identified in POMC fluid samples by 16S rRNA gene analysis. Species of Prevotella (91%), Neisseria (73%), Fusobacterium (73%), Porphyromonas (73%), and Propionibacterium (73%) were found to be highly prevalent in all patients. Streptococcus mitis (64%), Fusobacterium nucleatum (55%), Propionibacterium acnes (55%), Staphylococcus capitis (55%), and Streptococcus salivarius (55%) were detected in more than 6 of the 11 patients. The results obtained by the culture method were different from those obtained by 16S rRNA gene analysis, but both approaches may be necessary for the identification of pathogens, especially of bacteria that are difficult to detect by culture methods, and the development of rational treatments for patients with POMC.


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