scholarly journals Endosymbiont Communities in Pachyseris speciosa Highlight Geographical and Methodological Variations

2021 ◽  
Vol 8 ◽  
Author(s):  
Sudhanshi S. Jain ◽  
Lutfi Afiq-Rosli ◽  
Bar Feldman ◽  
Ismael Kunning ◽  
Oren Levy ◽  
...  
Keyword(s):  
Papua New Guinea ◽  
Internal Transcribed Spacer ◽  
High Throughput Sequencing ◽  
Quantitative Polymerase Chain Reaction ◽  
New Guinea ◽  
Contrast Variation ◽  
Chain Reaction ◽  
Local Conditions ◽  
Intraspecific Variations ◽  
Polymerase Chain

Reef-building corals live in symbiosis with the phototrophic dinoflagellate family Symbiodiniaceae, which comprises diverse genera such as Cladocopium and Durusdinium. Pachyseris speciosa, a widely distributed Indo-Pacific coral found in a variety of reef habitats, is known to be associated with these two Symbiodiniaceae genera, but little is known about the biogeographic variability of the endosymbiont communities across the region. In this study, the diversity and dominance patterns of Symbiodiniaceae at the western and eastern areas of the Central Indo-Pacific region were examined. We sampled Pachyseris speciosa colonies at seven and six sites in Singapore and Papua New Guinea, respectively, and genotyped their endosymbionts based on the internal transcribed spacer (ITS) markers using two distinct methods, quantitative polymerase chain reaction (qPCR) and high-throughput sequencing (HTS). Results showed 92% of all colonies in Singapore exhibiting Cladocopium dominance. There was a higher abundance of Durusdinium compared to Cladocopium in certain colonies from one site, Pulau Hantu (mean Durusdinium abundance of 90%, compared to 0–14% among all other sites). In contrast, variation in the endosymbiont communities was generally higher among sites in Papua New Guinea. Cladocopium expectedly dominated most colonies (75%), although colonies from Kimbe Bay (85%) and Kavieng (65%) showed Durusdinium dominance. Between localities, relative genus abundances based on qPCR were not significantly different, but HTS showed that the ratio of Durusdinium over Cladocopium was significantly higher in Papua New Guinea corals. Notably, 6% of colonies from Singapore and 15% from Papua New Guinea showed endosymbiont dominance patterns that were inconsistent between the two methods, underscoring the need for further validation of symbiotic algal quantification based on HTS. The richness of ITS2 type profiles was clearly lower among colonies from the impacted and turbid reefs of Singapore compared to the less urbanized reefs of Papua New Guinea. These coral intraspecific variations of Symbiodiniaceae communities within and among localities suggest that local conditions are important drivers of endosymbiosis and may ultimately influence corals’ resilience against global stressors such as ocean warming.

Metronidazole resistance in Trichomonas vaginalis from highland women in Papua New Guinea

Sexual Health ◽  
2009 ◽  
Vol 6 (4) ◽  
pp. 334 ◽  
Author(s):  
Jacqueline A. Upcroft ◽  
Linda A. Dunn ◽  
Tilda Wal ◽  
Sepehr Tabrizi ◽  
Maria G. Delgadillo-Correa ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Papua New Guinea ◽  
Trichomonas Vaginalis ◽  
Protozoan Parasite ◽  
New Guinea ◽  
Chain Reaction ◽  
Clinical Resistance ◽  
Polymerase Chain ◽  
High Level

Background: The prevalence of the sexually transmissible protozoan parasite Trichomonas vaginalis in the highlands of Papua New Guinea (PNG) has been reported to be as high as 46% and although not previously studied in Papua New Guinea, clinical resistance against metronidazole (Mz), the drug most commonly used to treat trichomoniasis, is well documented worldwide. This study was primarily aimed at assessing resistance to Mz in T. vaginalis strains from the Goroka region. Methods: Consenting patients presenting at the Goroka Base Hospital Sexually Transmitted Diseases (STD) Clinic and local women were asked to provide two vaginal swabs: one for culturing of the parasite; and one for polymerase chain reaction detection of T. vaginalis, Chlamydia trachomatis and Neisseria gonorrhoeae. T. vaginalis isolates were assayed for Mz susceptibility and a selection was genotyped. Results: The prevalence of T. vaginalis was determined to be 32.9% by culture and polymerase chain reaction of swabs among 82 local women and patients from the STD clinic. An unexpectedly high level of in vitro Mz resistance was determined with 17.4% of isolates displaying unexpectedly high resistance to Mz. The ability to identify isolates of T. vaginalis by genotyping was confirmed and the results revealed a more homogeneous T. vaginalis population in Papua New Guinea compared with isolates from elsewhere. Conclusion: T. vaginalis is highly prevalent in the Goroka region and in vitro Mz resistance data suggest that clinical resistance may become an issue.

AN IMPROVED METHOD FOR INHIBITOR FREE NUCLEIC ACIDS FROM POLYPHENOL RICH LEAVES OF MUSA SPP

2017 ◽  
Vol 23 (1) ◽  
Author(s):  
N.NANDHA KUMAR ◽  
K. SOURIANATHA SUNDARAM ◽  
D. SUDHAKAR ◽  
K.K. KUMAR
Keyword(s):  
Polymerase Chain Reaction ◽  
Quantitative Polymerase Chain Reaction ◽  
Proteinase K ◽  
Chain Reaction ◽  
Banana Plant ◽  
High Molecular Weight Dna ◽  
Musa Spp ◽  
Leaf Tissues ◽  
Polymerase Chain

Excessive presence of polysaccharides, polyphenol and secondary metabolites in banana plant affects the quality of DNA and it leads to difficult in isolating good quality of DNA. An optimized modified CTAB protocol for the isolation of high quality and quantity of DNA obtained from banana leaf tissues has been developed. In this protocol a slight increased salt (NaCl) concentration (2.0M) was used in the extraction buffer. Polyvinylpyrrolidone (PVP) and Octanol were used for the removal of polyphenols and polymerase chain reaction (PCR) inhibitors. Proteins like various enzymes were degraded by Proteinase K and removed by centrifugation from plant extract during the isolation process resulting in pure genomic DNA, ready to use in downstream applications including PCR, quantitative polymerase chain reaction (qPCR), ligation, restriction and sequencing. This protocol yielded a high molecular weight DNA isolated from polyphenols rich leaves of Musa spp which was free from contamination and colour. The average yields of total DNA from leaf ranged from 917.4 to 1860.9 ng/ìL. This modified CTAB protocol reported here is less time consuming 4-5h, reproducible and can be used for a broad spectrum of plant species which have polyphenol and polysaccharide compounds.

scholarly journals Analysis and validation of a highly sensitive one-step nested quantitative real-time polymerase chain reaction assay for specific detection of severe acute respiratory syndrome coronavirus 2

2020 ◽  
Vol 17 (1) ◽  
Author(s):  
Yang Zhang ◽  
Chunyang Dai ◽  
Huiyan Wang ◽  
Yong Gao ◽  
Tuantuan Li ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Real Time ◽  
Quantitative Polymerase Chain Reaction ◽  
Clinical Samples ◽  
Chain Reaction ◽  
Pcr Assay ◽  
Qrt Pcr ◽  
Highly Sensitive ◽  
One Step ◽  
Polymerase Chain

Abstract Background Coronavirus disease 2019 (COVID-19), caused by SARS-CoV-2, is posing a serious threat to global public health. Reverse transcriptase real-time quantitative polymerase chain reaction (qRT-PCR) is widely used as the gold standard for clinical detection of SARS-CoV-2. Due to technical limitations, the reported positive rates of qRT-PCR assay of throat swab samples vary from 30 to 60%. Therefore, the evaluation of alternative strategies to overcome the limitations of qRT-PCR is required. A previous study reported that one-step nested (OSN)-qRT-PCR revealed better suitability for detecting SARS-CoV-2. However, information on the analytical performance of OSN-qRT-PCR is insufficient. Method In this study, we aimed to analyze OSN-qRT-PCR by comparing it with droplet digital PCR (ddPCR) and qRT-PCR by using a dilution series of SARS-CoV-2 pseudoviral RNA and a quality assessment panel. The clinical performance of OSN-qRT-PCR was also validated and compared with ddPCR and qRT-PCR using specimens from COVID-19 patients. Result The limit of detection (copies/ml) of qRT-PCR, ddPCR, and OSN-qRT-PCR were 520.1 (95% CI: 363.23–1145.69) for ORF1ab and 528.1 (95% CI: 347.7–1248.7) for N, 401.8 (95% CI: 284.8–938.3) for ORF1ab and 336.8 (95% CI: 244.6–792.5) for N, and 194.74 (95% CI: 139.7–430.9) for ORF1ab and 189.1 (95% CI: 130.9–433.9) for N, respectively. Of the 34 clinical samples from COVID-19 patients, the positive rates of OSN-qRT-PCR, ddPCR, and qRT-PCR were 82.35% (28/34), 67.65% (23/34), and 58.82% (20/34), respectively. Conclusion In conclusion, the highly sensitive and specific OSN-qRT-PCR assay is superior to ddPCR and qRT-PCR assays, showing great potential as a technique for detection of SARS-CoV-2 in patients with low viral loads.

scholarly journals Modulation of the Tissue Expression Pattern of Zebrafish CRP-Like Molecules Suggests a Relevant Antiviral Role in Fish Skin

Biology ◽  
2021 ◽  
Vol 10 (2) ◽  
pp. 78
Author(s):  
Melissa Bello-Perez ◽  
Mikolaj Adamek ◽  
Julio Coll ◽  
Antonio Figueras ◽  
Beatriz Novoa ◽  
...  
Keyword(s):  
Gene Expression ◽  
Bacterial Infections ◽  
Quantitative Polymerase Chain Reaction ◽  
Tissue Expression ◽  
Interferon Response ◽  
Fish Skin ◽  
Tissue Expression Pattern ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Antiviral Role

Recent studies suggest that short pentraxins in fish might serve as biomarkers for not only bacterial infections, as in higher vertebrates including humans, but also for viral ones. These fish orthologs of mammalian short pentraxins are currently attracting interest because of their newly discovered antiviral activity. In the present work, the modulation of the gene expression of all zebrafish short pentraxins (CRP-like proteins, CRP1-7) was extensively analyzed by quantitative polymerase chain reaction. Initially, the tissue distribution of crp1-7 transcripts and how the transcripts varied in response to a bath infection with the spring viremia of carp virus, were determined. The expression of crp1-7 was widely distributed and generally increased after infection (mostly at 5 days post infection), except for crp1 (downregulated). Interestingly, several crp transcription levels significantly increased in skin. Further assays in mutant zebrafish of recombinant activation gene 1 (rag1) showed that all crps (except for crp2, downregulated) were already constitutively highly expressed in skin from rag1 knockouts and only increased moderately after viral infection. Similar results were obtained for most mx isoforms (a reporter gene of the interferon response), suggesting a general overcompensation of the innate immunity in the absence of the adaptive one.

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