Molecular Identification of Borrelia spp. from Ticks in Pastures nearby Livestock Farms in Korea

Ticks are vectors that spread pathogenic bacteria, viruses, and protozoa. As the number of ticks increases due to climate change, the importance of the study of tick-borne pathogens has also increased. This study was conducted to investigate the distribution of the major tick species causing Lyme borreliosis and regional differences in the prevalence of Borrelia spp. by tick species. Borrelia infection was confirmed not only in Ixodes ticks, which are the major vectors of Borrelia spp., but also in Haemaphysalis and Amblyomma ticks. PCR targeting the 5S-23S rRNA intergenic spacer region (rrf-rrl) was performed to confirm Borrelia positivity. A total of 6102 ticks (736 pools) were tested, and the proportion was Haemaphysalis longicornis nymphs and adults at 69.2%, Haemaphysalis flava nymphs and adults at 13.9%, Haemaphysalis spp. larva at 14.3%, Ixodes nipponensis at 0.8%, and Amblyomma testudinarium at 1.9%. Ixodes nipponensis showed the highest minimum infection rate (MIR: 34.00; 17 pools/50 ticks) for Borrelia spp., followed by A. testudinarium (MIR: 0.88), and H. longicornis (MIR: 0.05). In particular, to our knowledge Borrelia infection was first confirmed in A. testudinarium in Korea. As a result of phylogenetic analysis, all sequences were grouped with Borreliaafzelii isolates and showed a close relationship with high identity. Considering that B. afzelii causes infectious zoonotic diseases, continuous monitoring and attention are needed, although it has a low prevalence in this study.

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Molecular Detection and Characterization of Borrelia garinii (Spirochaetales: Borreliaceae) in Ixodes nipponensis (Ixodida: Ixodidae) Parasitizing a Dog in Korea

Pathogens ◽

10.3390/pathogens8040289 ◽

2019 ◽

Vol 8 (4) ◽

pp. 289 ◽

Cited By ~ 2

Author(s):

Seung-Hun Lee ◽

Youn-Kyoung Goo ◽

Paul John L. Geraldino ◽

Oh-Deog Kwon ◽

Dongmi Kwak

Keyword(s):

Molecular Detection ◽

Single Gene ◽

Protein A ◽

Intergenic Spacer ◽

Surface Protein ◽

23S Rrna ◽

Borrelia Garinii ◽

Intergenic Spacer Region ◽

Individual Level ◽

Pcr Targeting

The present study aimed to detect and characterize Borrelia spp. in ticks attached to dogs in Korea. Overall, 562 ticks (276 pools) attached to dogs were collected and tested for Borrelia infection by PCR targeting the 5S-23S rRNA intergenic spacer region (rrf-rrl). One tick larva (pool level, 0.4%; individual level, 0.2%) was confirmed by sequencing Borrelia garinii, a zoonotic pathogen. For molecular characterization, the outer surface protein A (ospA) and flagellin genes were analyzed. Phylogenetic ospA analysis distinguished B. garinii from B. bavariensis, which has been recently identified as a novel Borrelia species. On the other hand, phylogenetic analysis showed that single gene analysis involving rrf-rrl or flagellin was not sufficient to differentiate B. garinii from B. bavariensis. In addition, the B. garinii-infected tick was identified as Ixodes nipponensis by sequencing according to mitochondrial 16S rRNA and the second transcribed spacer region. To our knowledge, this is the first study to report the molecular detection of B. garinii in I. nipponensis parasitizing a dog in Korea. Continuous monitoring of tick-borne pathogens in ticks attached to animals is required to avoid disease distribution and possible transmission to humans.

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Prevalence of Lyme Disease Borrelia spp. in Ticks from Migratory Birds on the Japanese Mainland

Applied and Environmental Microbiology ◽

10.1128/aem.66.3.982-986.2000 ◽

2000 ◽

Vol 66 (3) ◽

pp. 982-986 ◽

Cited By ~ 46

Author(s):

Fubito Ishiguro ◽

Nobuhiro Takada ◽

Toshiyuki Masuzawa ◽

Takako Fukui

Keyword(s):

Lyme Disease ◽

Migratory Birds ◽

Intergenic Spacer ◽

23S Rrna ◽

Polymorphism Analysis ◽

Ixodes Persulcatus ◽

Haemaphysalis Longicornis ◽

Borrelia Garinii ◽

Haemaphysalis Flava ◽

Large Animals

ABSTRACT Borrelia sp. prevalence in ticks on migratory birds was surveyed in central Japan. In autumn, a total of 1,733 birds representing 40 species were examined for ticks. A total of 361 ticks were obtained from 173 birds of 15 species, and these ticks were immature Haemaphysalis flava (94.4%), Haemaphysalis longicornis, Ixodes columnae, Ixodes persulcatus, Ixodes turdus, and an unidentifiedIxodes species. Of these, 27 juveniles of H. flava on Turdus pallidus, Turdus cardis, or Emberiza spodocephala, 2 juveniles of I. persulcatus on T. pallidus, and 1 female H. flava molted from a T. pallidus-derived nymph were positive for the presence of Borrelia by Barbour-Stoenner-Kelly culture passages. In spring, a total of 16 ticks obtained from 102 birds of 21 species were negative for the spirochete. Isolates from 15 ticks were characterized by 5S-23S rRNA intergenic spacer restriction fragment length polymorphism analysis; all isolates were identified as Borrelia garinii with pattern B/B′ based on the previous patterning. According to the intergenic spacer sequences, 2 of 15 isolates, strains Fi14f and Fi24f, were highly similar to B. garinii strains 935T of Korea and ChY13p of Inner Mongolia, China, respectively. These findings indicate that Lyme disease-causing B. garinii may have been introduced to Japan by migratory birds from northeastern China via Korea. Additionally, a case of transstadial transmission of B. garinii from nymph to adult H. flava suggests that the infected H. flava may transmit Borrelia to large animals.

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Tick species from cattle in the Adama Region of Ethiopia and pathogens detected

Experimental and Applied Acarology ◽

10.1007/s10493-021-00623-5 ◽

2021 ◽

Author(s):

Tafese Beyene Tufa ◽

Silke Wölfel ◽

Dana Zubriková ◽

Bronislava Víchová ◽

Martin Andersson ◽

...

Keyword(s):

Disease Transmission ◽

Tick Species ◽

Pathogenic Bacteria ◽

Intergenic Spacer ◽

Farm Animals ◽

Ehrlichia Ruminantium ◽

Intergenic Spacer Region ◽

Dna Yield ◽

Rhipicephalus Pulchellus ◽

Tick Borne Disease

AbstractTicks will diminish productivity among farm animals and transmit zoonotic diseases. We conducted a study to identify tick species infesting slaughter bulls from Adama City and to screen them for tick-borne pathogens. In 2016, 291 ticks were collected from 37 bulls in Adama, which were ready for slaughter. Ticks were identified morphologically. Total genomic DNA was extracted from ticks and used to test for Rickettsia spp. with real-time PCR. Species identification was done by phylogenetic analysis using sequencing that targeted the 23S-5S intergenic spacer region and ompA genes. Four tick species from two genera, Amblyomma and Rhipicephalus, were identified. Amblyomma cohaerens was the dominant species (n = 241, 82.8%), followed by Amblyomma variegatum (n = 22, 7.5%), Rhipicephalus pulchellus (n = 19, 6.5%), and Rhipicephalus decoloratus (n = 9, 3.0%). Among all ticks, 32 (11%) were positive for Rickettsia spp. and 15 (5.2%) of these were identified as R. africae comprising at least two genetic clades, occurring in A. variegatum (n = 10) and A. cohaerens (n = 5). The remainder of Rickettsia-positive samples could not be amplified due to low DNA yield. Furthermore, another 15 (5.2%) samples carried other pathogenic bacteria: Ehrlichia ruminantium (n = 9; 3.1%) in A. cohaerens, Ehrlichia sp. (n = 3; 1%) in Rh. pulchellus and A. cohaerens, Anaplasma sp. (n = 1; 0.5%) in A. cohaerens, and Neoehrlichia mikurensis (n = 2; 0.7%) in A. cohaerens. All ticks were negative for Bartonella spp., Babesia spp., Theileria spp., and Hepatozoon spp. We reported for the first time E. ruminatium, N. mikurensis, Ehrlichia sp., and Anaplasma sp. in A. cohaerens. Medically and veterinarily important pathogens were mostly detected from A. variegatum and A. cohaerens. These data are relevant for a One-health approach for monitoring and prevention of tick-borne disease transmission.

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Diversity of Salmonella Strains Isolated from the Aquatic Environment as Determined by Serotyping and Amplification of the Ribosomal DNA Spacer Regions

Applied and Environmental Microbiology ◽

10.1128/aem.66.4.1544-1552.2000 ◽

2000 ◽

Vol 66 (4) ◽

pp. 1544-1552 ◽

Cited By ~ 117

Author(s):

Julia Baudart ◽

Karine Lemarchand ◽

Anne Brisabois ◽

Philippe Lebaron

Keyword(s):

Pathogenic Bacteria ◽

Natural Waters ◽

Pcr Amplification ◽

Intergenic Spacer ◽

Rrna Genes ◽

23S Rrna ◽

Natural Environments ◽

Storm Events ◽

Intergenic Spacer Region ◽

Coastal Watershed

ABSTRACT Salmonella species are pathogenic bacteria often detected in sewage, freshwater, marine coastal water, and groundwater.Salmonella spp. can survive for long periods in natural waters, and the persistence of specific and epidemic strains is of great concern in public health. However, the diversity of species found in the natural environment remains unknown. The aim of this study was to investigate the diversity of Salmonellastrains isolated from different natural aquatic systems within a Mediterranean coastal watershed (river, wastewater, and marine coastal areas). A total of 574 strains isolated from these natural environments were identified by both conventional serotyping and the ribosomal spacer-heteroduplex polymorphism (RS-HP) method (M. A. Jensen and N. Straus, PCR Methods Appl. 3:186–194, 1993). More than 40 different serotypes were found, and some serotypes probably mobilized from widespread animal-rearing activities were detected only during storm events. These serotypes may be good indicators of specific contamination sources. Furthermore, the RS-HP method based on the PCR amplification of the intergenic spacer region between the 16S and 23S rRNA genes can produce amplicon profiles allowing the discrimination of species at both serotype and intraserotype levels. This method represents a powerful tool that could be used for rapid typing of Salmonella isolates.

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Longitudinal monitoring of the dynamics of infections due to Bartonella species in UK woodland rodents

Epidemiology and Infection ◽

10.1017/s095026880100526x ◽

2001 ◽

Vol 126 (2) ◽

pp. 323-329 ◽

Cited By ~ 60

Author(s):

R. J. BIRTLES ◽

S. M. HAZEL ◽

M. BENNETT ◽

K. BOWN ◽

D. RAOULT ◽

...

Keyword(s):

Sequence Comparison ◽

Intergenic Spacer ◽

23S Rrna ◽

Enzyme Analysis ◽

Restriction Enzyme Analysis ◽

Bartonella Species ◽

Intergenic Spacer Region ◽

Specific Pcr ◽

Time Specific ◽

Pcr Targeting

Blood samples were repeatedly collected from 12 sympatric woodland rodents over a 12-month period and DNA extracts from each were incorporated into a bartonella-specific PCR targeting a fragment of the 16S/23S rRNA intergenic spacer region (ISR). The composition of each amplicon was analysed using restriction enzyme analysis (REA) and base sequence comparison. Bartonella DNA was detected in 70 of 109 samples. Eleven samples contained DNA derived from more than one strain. Sequence analysis of 62 samples found 12 sequence variants (ISR genotypes) that were provisionally assigned to 5 different species, 2 of which were newly recognized. Up to five different species were detected in each animal. On about two-thirds of occasions, a species detected 1 month was not there the next, but never was a genotype superseded by another of the same species. However, a genotype could be re-encountered months later in the same animal, even if interim samples contained other genotypes. Our results suggest that although most animals are bacteraemic most of the time, specific infections are often superseded and that a complex and dynamic epidemiology of bartonella bacteraemias exists in woodland rodents.

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Genomic fingerprinting ofFrankiamicrosymbionts fromCeanothuscopopulations using repetitive sequences and polymerase chain reactions

Canadian Journal of Botany ◽

10.1139/b99-069 ◽

1999 ◽

Vol 77 (9) ◽

pp. 1220-1230 ◽

Cited By ~ 3

Author(s):

Soon-Chun Jeong ◽

David D Myrold

Keyword(s):

Dna Sequencing ◽

Dna Analysis ◽

Repetitive Sequences ◽

Intergenic Spacer ◽

Rrna Genes ◽

23S Rrna ◽

Genomic Fingerprinting ◽

Chain Reactions ◽

Intergenic Spacer Region ◽

Polymerase Chain

Specificity between Ceanothus species and their microsymbionts, Frankia, were investigated with nodules collected from three geographically separated copopulations of Ceanothus species. Nodules were analyzed using DNA sequencing and repetitive sequence polymerase chain reaction (rep-PCR) techniques. DNA sequencing of the intergenic spacer region between 16S and 23S rRNA genes suggested that Ceanothus-microsymbiotic Frankia are closely related at the intraspecific level. Diversity of the microsymbionts was further analyzed by genomic fingerprinting using repetitive sequences and PCR. A newly designed direct repeat (DR) sequence and a BOX sequence were used as PCR primers after justification that these primers can generate Frankia-specific fingerprints from nodule DNA. Analysis of the nodules using BOX- and DR-PCR showed that Ceanothus-microsymbiotic Frankia exhibited less diversity within each copopulation than among copopulations. These data suggested that geographic separation plays a more important role for divergence of Ceanothus-microsymbiotic Frankia than host plant.Key words: Frankia, Ceanothus, rep-PCR, diversity.

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Differentiation of group VIII Spiroplasma strains with sequences of the 16S–23S rDNA intergenic spacer region

Canadian Journal of Microbiology ◽

10.1139/w04-091 ◽

2004 ◽

Vol 50 (12) ◽

pp. 1061-1067 ◽

Cited By ~ 14

Author(s):

Laura B Regassa ◽

Kimberly M Stewart ◽

April C Murphy ◽

Frank E French ◽

Tao Lin ◽

...

Keyword(s):

Intergenic Spacer ◽

Analytical Models ◽

23S Rrna ◽

Rrna Gene ◽

Group Viii ◽

Similarity Coefficients ◽

Intergenic Spacer Region ◽

Intergenic Sequences ◽

The Stability ◽

The 16S Rrna Gene

Spiroplasma species (Mollicutes: Spiroplasmataceae) are associated with a wide variety of insects, and serology has classified this genus into 34 groups, 3 with subgroups. The 16S rRNA gene has been used for phylogenetic analysis of spiroplasmas, but this approach is uninformative for group VIII because the serologically distinct subgroups generally have similarity coefficients >0.990. Therefore, we investigated the utility of the 16S–23S rRNA spacer region as a means to differentiate closely related subgroups or strains. We generated intergenic sequences and detailed serological profiles for 8 group VIII Spiroplasma strains. Sequence analyses using Maximum Parsimony, Neighbor Joining, and Maximum Likelihood placed the strains into 2 clades. One clade consisted of strains BARC 2649 and GSU5367. The other clade was divided into clusters containing representatives of the 3 designated group VIII subgroups (EA-1, DF-1, and TAAS-1) and 3 previously unclassified strains. The stability of the positions of the strains in various analytical models and the ability to provide robust support for groupings tentatively supported by serology indicates that the 16S–23S intergenic rDNA sequence will prove useful in intragroup analysis of group VIII spiroplasmas.Key words: Mollicutes, Spiroplasma, phylogeny, Tabanidae.

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A putative transposase gene in the 16S–23S rRNA intergenic spacer region of Mycoplasma imitans

Microbiology ◽

10.1099/mic.0.26629-0 ◽

2004 ◽

Vol 150 (4) ◽

pp. 1023-1029 ◽

Cited By ~ 28

Author(s):

Ryô Harasawa ◽

David G. Pitcher ◽

Ana S. Ramírez ◽

Janet M. Bradbury

Keyword(s):

Relative Size ◽

Intergenic Spacer ◽

Its Region ◽

23S Rrna ◽

Mycoplasma Species ◽

Insertion Element ◽

Transposase Gene ◽

Intergenic Spacer Region ◽

Pcr Products ◽

Type Strains

Examination of the nucleotide sequences of the 16S–23S intergenic transcribed spacer (ITS) region of Mycoplasma imitans and Mycoplasma gallisepticum identified a putative transposase gene located only in the ITS of M. imitans, which can be used as a genetic marker to distinguish these two species. The relative size of the PCR products of the ITS region allowed a clear distinction to be made between strains of M. imitans and M. gallisepticum, both of which could be readily discriminated from the type strains of all the other recognized avian Mycoplasma species. In addition, the putative transposase gene assigned in the ITS of M. imitans was shown to include a sequence homologous to that of the P75 gene of M. gallisepticum. This is believed to be the first description of an insertion element in the rRNA operon region of a mycoplasma species.

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