scholarly journals Fluorescence in situ hybridization in species-specific diagnosis of ovine Campylobacter abortions

2020 ◽  
Vol 32 (3) ◽  
pp. 413-419
Author(s):  
Godelind A. Wolf-Jäckel ◽  
Mette Boye ◽  
Øystein Angen ◽  
Matthias Müller ◽  
Tim K. Jensen
Keyword(s):  
In Situ Hybridization ◽  
Fluorescence In Situ Hybridization ◽  
Pathogenic Bacteria ◽  
Bacterial Species ◽  
Detection Tool ◽  
Major Species ◽  
Species Specific ◽  
Campylobacter Infection ◽  
Culture Negative

Campylobacter infection is a leading cause of ovine abortion worldwide. Campylobacter fetus and C. jejuni are the major species involved. We report herein on abortion storms in 4 Danish sheep flocks. Initially, no pathogenic bacteria were isolated from placental and fetal tissues on aerobic and selective media despite the presence of severe suppurative and necrotizing placentitis with numerous bacteria located intracellularly in trophoblasts. Fluorescence in situ hybridization (FISH) was then applied on abortion material from 13 cases; species-specific oligonucleotide probes directed against either C. fetus or C. jejuni were used in combination with a general bacterial probe. C. fetus was detected as the only lesion-associated bacterial species in 4 cases from 2 flocks, and C. jejuni in 6 cases from the other 2 flocks, thereby establishing the likely etiology of the abortion storms in all 4 flocks. FISH is a useful detection tool in culture-negative cases with tissue lesions suggestive of bacterial infection. Furthermore, FISH is a fast and economical method to detect and identify the zoonotic agent Campylobacter within ovine abortion material.

Detection of Bacteria by Fluorescence in Situ Hybridization in Culture-Negative Soft Tissue Filler Lesions

2009 ◽  
Vol 35 ◽  
pp. 1620-1624 ◽  
Author(s):  
THOMAS BJARNSHOLT ◽  
TIM TOLKER-NIELSEN ◽  
MICHAEL GIVSKOV ◽  
MARTIN JANSSEN ◽  
LISE HANNE CHRISTENSEN
Keyword(s):  
In Situ Hybridization ◽  
Soft Tissue ◽  
Fluorescence In Situ Hybridization ◽  
Soft Tissue Filler ◽  
Culture Negative

scholarly journals Comparative study of four Mystus species (Bagridae, Siluriformes) from Thailand: insights into their karyotypic diversity

2021 ◽  
Vol 15 (2) ◽  
pp. 119-136
Author(s):  
Pun Yeesin ◽  
Phichaya Buasriyot ◽  
Sukhonthip Ditcharoen ◽  
Patcharaporn Chaiyasan ◽  
Chatmongkon Suwannapoom ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Comparative Study ◽  
Fluorescence In Situ Hybridization ◽  
Chromosome Numbers ◽  
Single Pair ◽  
Molecular Cytogenetic ◽  
Karyotypic Diversity ◽  
Species Specific

Karyotypes of four catfishes of the genus Mystus Scopoli, 1777 (family Bagridae), M. atrifasciatus Fowler, 1937, M. mysticetus Roberts, 1992, M. singaringan (Bleeker, 1846) and M. wolffii (Bleeker, 1851), were analysed by conventional and Ag-NOR banding as well as fluorescence in situ hybridization (FISH) techniques. Microsatellite d(GC)15, d(CAA)10, d(CAT)10 and d(GAA)10 repeat probes were applied in FISH. The obtained data revealed that the four studied species have different chromosome complements. The diploid chromosome numbers (2n) and the fundamental numbers (NF) range between 52 and 102, 54 and 104, 56 and 98, or 58 and 108 in M. mysticetus, M. atrifasciatus, M. singaringan or M. wolffii, respectively. Karyotype formulae of M. mysticetus, M. atrifasciatus, M. singaringan and M. wolffii are 24m+26sm+4a, 26m+24sm+2a, 24m+18sm+14a and 30m+22sm+6a, respectively. A single pair of NORs was identified adjacent to the telomeres of the short arm of chromosome pairs 3 (metacentric) in M. atrifasciatus, 20 (submetacentric) in M. mysticetus, 15 (submetacentric) in M. singaringan, and 5 (metacentric) in M. wolffii. The d(GC)15, d(CAA)10, d(CAT)10 and d(GAA)10 repeats were abundantly distributed in species-specific patterns. Overall, we present a comparison of cytogenetic and molecular cytogenetic patterns of four species from genus Mystus providing insights into their karyotype diversity in the genus.

scholarly journals Comparative study of four Mystus species (Bagridae, Siluriformes) from Thailand: insights into their karyotypic diversity

2021 ◽  
Vol 15 (2) ◽  
pp. 119-136
Author(s):  
Pun Yeesin ◽  
Phichaya Buasriyot ◽  
Sukhonthip Ditcharoen ◽  
Patcharaporn Chaiyasan ◽  
Chatmongkon Suwannapoom ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Comparative Study ◽  
Fluorescence In Situ Hybridization ◽  
Chromosome Numbers ◽  
Single Pair ◽  
Molecular Cytogenetic ◽  
Karyotypic Diversity ◽  
Species Specific

Karyotypes of four catfishes of the genus Mystus Scopoli, 1777 (family Bagridae), M. atrifasciatus Fowler, 1937, M. mysticetus Roberts, 1992, M. singaringan (Bleeker, 1846) and M. wolffii (Bleeker, 1851), were analysed by conventional and Ag-NOR banding as well as fluorescence in situ hybridization (FISH) techniques. Microsatellite d(GC)15, d(CAA)10, d(CAT)10 and d(GAA)10 repeat probes were applied in FISH. The obtained data revealed that the four studied species have different chromosome complements. The diploid chromosome numbers (2n) and the fundamental numbers (NF) range between 52 and 102, 54 and 104, 56 and 98, or 58 and 108 in M. mysticetus, M. atrifasciatus, M. singaringan or M. wolffii, respectively. Karyotype formulae of M. mysticetus, M. atrifasciatus, M. singaringan and M. wolffii are 24m+26sm+4a, 26m+24sm+2a, 24m+18sm+14a and 30m+22sm+6a, respectively. A single pair of NORs was identified adjacent to the telomeres of the short arm of chromosome pairs 3 (metacentric) in M. atrifasciatus, 20 (submetacentric) in M. mysticetus, 15 (submetacentric) in M. singaringan, and 5 (metacentric) in M. wolffii. The d(GC)15, d(CAA)10, d(CAT)10 and d(GAA)10 repeats were abundantly distributed in species-specific patterns. Overall, we present a comparison of cytogenetic and molecular cytogenetic patterns of four species from genus Mystus providing insights into their karyotype diversity in the genus.

Physical localisation of repetitive DNA sequences in Alstroemeria: karyotyping of two species with species-specific and ribosomal DNA

Genome ◽  
1997 ◽  
Vol 40 (5) ◽  
pp. 652-658 ◽  
Author(s):  
Silvan A. Kamstra ◽  
Anja G. J. Kuipers ◽  
Marjo J. De Jeu ◽  
M. S. Ramanna ◽  
Evert Jacobsen
Keyword(s):  
In Situ Hybridization ◽  
Fluorescence In Situ Hybridization ◽  
Repetitive Dna ◽  
Ribosomal Dna ◽  
Dna Sequences ◽  
Repetitive Dna Sequences ◽  
Metaphase Chromosomes ◽  
Rdna Sites ◽  
Species Specific

Fluorescence in situ hybridization (FISH) was used to localise two species-specific repetitive DNA sequences, A001-I and D32-13, and two highly conserved 25S and 5S rDNA sequences on the metaphase chromosomes of two species of Alstroemeria. The Chilean species, Alstroemeria aurea (2n = 16), has abundant constitutive heterochromatin, whereas the Brazilian species, Alstroemeria inodora, has hardly any heterochromatin. The A. aurea specific A001-I probe hybridized specifically to the C-band regions on all chromosomes. The FISH patterns on A. inodora chromosomes using species-specific probe D32–13 resembled the C-banding pattern and the A001-I pattern on A. aurea chromosomes. There were notable differences in number and distribution of rDNA sites between the two species. The 25S rDNA probe revealed 16 sites in A. aurea that closely colocalised with A001-I sites and 12 in A. inodora that were predominantly detected in the centromeric regions. FISH karyotypes of the two Alstroemeria species were constructed accordingly, enabling full identification of all individual chromosomes. These FISH karyotypes will be useful for monitoring the chromosomes of both Alstroemeria species in hybrids and backcross derivatives.Key words: Alstroemeria, fluorescence in situ hybridization, FISH, repetitive DNA, ribosomal DNA, karyotype.

scholarly journals Development of a fixation-free fluorescence in situ hybridization for the detection of Salmonella species

2020 ◽  
Vol 5 (1) ◽  
Author(s):  
Oluwawemimo Adebowale ◽  
Liam Good
Keyword(s):  
In Situ Hybridization ◽  
Fluorescence In Situ Hybridization ◽  
Effective Control ◽  
Cellular Delivery ◽  
Early Management ◽  
Fish Method ◽  
Severe Gastroenteritis ◽  
Species Specific ◽  
Single Reaction

Abstract Salmonella is one of the most important infectious bacteria causing severe gastroenteritis and deaths in humans and animals, and the prompt diagnosis is crucial for effective control and treatment. The detection of Salmonella still depends principally on culture-based methods, which is time-consuming and laborious. Recently, polyhexamethylene biguanide (PHMB) was discovered to have cellular delivery properties and its combination with the fluorescence in situ hybridization (FISH) method was exploited for oligomer delivery and the rapid detection of Salmonella spps in this study. Cell-associated fluorescence was quantified using Volocity® 3-D image analysis software (Volocity 6.3, PerkinElmer, Inc.). PHMB complexed with fluorophore—labelled species-specific oligomers permeabilized freshly grown viable strains of Salmonella cells and mediated strong cell-associated fluorescence signals. This strategy further enabled a fixation-free protocol and hybridization in a single reaction. Using the modified FISH method, monoculture Salmonella strains were validated as well as detected in artificially contaminated water and milk within a turnaround period of 5 h. The method was observed to be comparable with the standard FISH technique (sFISH; P > 0.05). The findings suggest that fixation-free delivery and hybridization of oligomers using PHMB can provide a simplified and prompt strategy for Salmonella detection at the species level, and promote early management responses to the disease and appropriate antimicrobial therapy.

scholarly journals Large-Scale Species-Specific Microbial Identification by Fluorescence In Situ Hybridization

2020 ◽  
Vol 118 (3) ◽  
pp. 464a
Author(s):  
Sungho Kim ◽  
Jae-Kyeong Im ◽  
Seungmin Yun ◽  
Hwasooo Koh ◽  
Donghoon Kang ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Fluorescence In Situ Hybridization ◽  
Large Scale ◽  
Microbial Identification ◽  
Species Specific

scholarly journals Identification of Dekkera bruxellensis(Brettanomyces) from Wine by Fluorescence In Situ Hybridization Using Peptide Nucleic Acid Probes

2001 ◽  
Vol 67 (2) ◽  
pp. 938-941 ◽  
Author(s):  
Henrik Stender ◽  
Cletus Kurtzman ◽  
Jens J. Hyldig-Nielsen ◽  
Ditte Sørensen ◽  
Adam Broomer ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Nucleic Acid ◽  
Fluorescence In Situ Hybridization ◽  
Peptide Nucleic Acid ◽  
Dekkera Bruxellensis ◽  
Specific Sequence ◽  
Nucleic Acid Probes ◽  
Yeast Strains ◽  
Species Specific

ABSTRACT A new fluorescence in situ hybridization method using peptide nucleic acid (PNA) probes for identification ofBrettanomyces is described. The test is based on fluorescein-labeled PNA probes targeting a species-specific sequence of the rRNA of Dekkera bruxellensis. The PNA probes were applied to smears of colonies, and results were interpreted by fluorescence microscopy. The results obtained from testing 127 different yeast strains, including 78 Brettanomycesisolates from wine, show that the spoilage organismBrettanomyces belongs to the species D. bruxellensis and that the new method is able to identifyBrettanomyces (D. bruxellensis) with 100% sensitivity and 100% specificity.

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