Discriminating Lineages of Batrachochytrium dendrobatidis using quantitative PCR

AbstractThe ability to detect and monitor infectious disease in a phylogenetically informative manner is critical for their management. Phylogenetically informative diagnostic tests enable patterns of pathogen introduction or changes in the distribution of genotypes to be measured, enabling research into the ecology of the pathogen. Batrachochytrium dendrobatidis (Bd), a causative agent of chytridiomycosis in amphibian populations, emerged worldwide in the 21st century and is composed of six lineages which are display varying levels of virulence in their hosts.Research into the distribution, ecology and pathogenicity of these lineages has been hampered by an inability to type lineage efficiently. Here, we describe a lineage-specific TaqMan qPCR assay that differentiates the two lineages of Bd most commonly associated with chytridiomycosis: BdGPL and BdCAPE. We demonstrate how this assay can be used for the surveillance of wild populations of amphibians in Southern Africa using skin swabs, tissue samples and cultured isolates.

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TaqMan qPCR assay for detecting Batrachochytrium dendrobatidis (Bd) v1 (protocols.io.bn2zmgf6)

protocols.io ◽

10.17504/protocols.io.bn2zmgf6 ◽

2020 ◽

Author(s):

Omneya Ahmed ◽

Mats T ◽

Tomas Larsson ◽

Alexander Eiler

Keyword(s):

Batrachochytrium Dendrobatidis ◽

Qpcr Assay ◽

Taqman Qpcr Assay ◽

Taqman Qpcr

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Batrachochytrium dendrobatidis is present in Poland and associated with reduced fitness in wild populations of Pelophylax lessonae

Diseases of Aquatic Organisms ◽

10.3354/dao03121 ◽

2017 ◽

Vol 124 (3) ◽

pp. 241-245

Author(s):

K Kolenda ◽

A Najbar ◽

M Ogielska ◽

V Baláž

Keyword(s):

Batrachochytrium Dendrobatidis ◽

Wild Populations

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Evidence for the Use of Mucus Swabs to Detect Renibacterium salmoninarum in Brook Trout

Pathogens ◽

10.3390/pathogens10040460 ◽

2021 ◽

Vol 10 (4) ◽

pp. 460

Author(s):

Tawni B. Riepe ◽

Victoria Vincent ◽

Vicki Milano ◽

Eric R. Fetherman ◽

Dana L. Winkelman

Keyword(s):

Brook Trout ◽

Sampling Methods ◽

Kidney Tissue ◽

Detection Methods ◽

Fish Health ◽

Wild Populations ◽

Renibacterium Salmoninarum ◽

Tissue Samples ◽

Future Studies ◽

Aquaculture Facilities

Efforts to advance fish health diagnostics have been highlighted in many studies to improve the detection of pathogens in aquaculture facilities and wild fish populations. Typically, the detection of a pathogen has required sacrificing fish; however, many hatcheries have valuable and sometimes irreplaceable broodstocks, and lethal sampling is undesirable. Therefore, the development of non-lethal detection methods is a high priority. The goal of our study was to compare non-lethal sampling methods with standardized lethal kidney tissue sampling that is used to detect Renibacterium salmoninarum infections in salmonids. We collected anal, buccal, and mucus swabs (non-lethal qPCR) and kidney tissue samples (lethal DFAT) from 72 adult brook trout (Salvelinus fontinalis) reared at the Colorado Parks and Wildlife Pitkin Brood Unit and tested each sample to assess R. salmoninarum infections. Standard kidney tissue detected R. salmoninarum 1.59 times more often than mucus swabs, compared to 10.43 and 13.16 times more often than buccal or anal swabs, respectively, indicating mucus swabs were the most effective and may be a useful non-lethal method. Our study highlights the potential of non-lethal mucus swabs to sample for R. salmoninarum and suggests future studies are needed to refine this technique for use in aquaculture facilities and wild populations of inland salmonids.

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Spotlight on PSMA as a new theranostic biomarker for bladder cancer

Scientific Reports ◽

10.1038/s41598-021-89160-0 ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Maria Maddalena Tumedei ◽

Sara Ravaioli ◽

Federica Matteucci ◽

Monica Celli ◽

Ugo De Giorgi ◽

...

Keyword(s):

Bladder Cancer ◽

Diagnostic Tests ◽

Radionuclide Therapy ◽

Tumor Type ◽

Prostate Specific Membrane Antigen ◽

Bladder Tissue ◽

Tissue Samples ◽

Specific Membrane ◽

Psma Expression ◽

Degree Of Extension

AbstractBladder cancer (BCa) patients are diagnosed by cytology and cystoscopy. However, these diagnostic tests bear some limitations. We sought for reliable biomarkers to better determine BCa extension. Prostate-specific membrane antigen (PSMA) appears to fulfill this requirement in prostate cancer but its role in BCa has not been established yet. We then analyzed 87 bladder tissue samples from 74 patients assessing PSMA expression by immunohistochemistry. The median PSMA expression, exclusively found in tumor neovasculature, in terms of H-score significantly differed between non-tumor samples and tumor samples (p = 0.00288) showing a higher neovasculature-related PSMA expression. No differences were observed in relation to tumor type, grade and stage. BCa neovasculature-related PSMA overexpression may be useful in defining the degree of extension of the neoplasm. In addition, testing PSMA expression by immunohistochemistry may hold theranostic implications both considering anti-angiogenesis agents and radio-labelled PSMA ligands for intracavitary radionuclide therapy. In our opinion, BCa neovasculature-related PSMA overexpression may be considered an apt target for anti-angiogenesis and radionuclide treatment in BCa, once the evaluation of tumor-retention time for the appropriateness of long half-life therapeutic PSMA ligands as radionuclide treatment will be performed.

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Assessment of copper accumulation in archived liver specimens from cats

Journal of Feline Medicine and Surgery ◽

10.1177/1098612x20961350 ◽

2020 ◽

pp. 1098612X2096135

Author(s):

Punyamanee Yamkate ◽

Randi M Gold ◽

Panagiotis G Xenoulis ◽

Katja Steiger ◽

David C Twedt ◽

...

Keyword(s):

Infectious Disease ◽

Hepatic Steatosis ◽

Dry Weight ◽

Reference Interval ◽

Copper Accumulation ◽

Zonal Distribution ◽

Tissue Samples ◽

Hepatic Copper ◽

Liver Copper ◽

Flame Atomic Absorption

Objectives The aim of this study was to assess hepatic copper concentrations and zonal distribution in cat liver specimens. Methods For this study, 121 archived, formalin-fixed, paraffin-embedded liver specimens from cats were used. Tissue sections were stained for copper with rhodanine and scored from 0 (no copper accumulation) to 5 (panlobular copper accumulation). The tissue specimens were then deparaffinized and hepatic copper concentrations were measured using flame atomic absorption spectroscopy. Results Tissue samples were categorized into four groups based on histopathologic findings: (1) no significant histopathologic hepatic changes (n = 66); (2) hepatic steatosis (n = 18); (3) inflammatory or infectious disease (n = 24); and (4) neoplasia (n = 13). Of the 121 specimens, 13 (11%) stained positive for copper, with three having a score ⩾3. Thirty-seven specimens (31%) had copper concentrations above the reference interval ([RI] <180 µg/g dry weight liver). Copper concentrations in cats with hepatic inflammatory or infectious disease were significantly higher than cats with hepatic steatosis ( P = 0.03). Copper-staining score and concentration were positively correlated ( rs = 0.46, P <0.001). Conclusions and relevance Despite the fact that 31% of specimens had copper concentrations above the RI, only 11% showed positive copper staining and only 2.5% had a score ⩾3. Our findings suggest that hepatic copper concentrations greater than the upper limit of the RI are relatively common in cats. Further studies to determine the factors that influence hepatic copper staining in cats and to establish contemporary RIs for hepatic copper in healthy cats are warranted.

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Dynamics of Specific cfDNA Fragments in the Plasma of Full Marathon Participants

Genes ◽

10.3390/genes12050676 ◽

2021 ◽

Vol 12 (5) ◽

pp. 676

Author(s):

Takehito Sugasawa ◽

Shin-ichiro Fujita ◽

Tomoaki Kuji ◽

Noriyo Ishibashi ◽

Kenshirou Tamai ◽

...

Keyword(s):

Exercise Physiology ◽

High Sensitivity ◽

Genomic Region ◽

Qpcr Assay ◽

Strongly Correlated ◽

Healthy Humans ◽

Blood Cell And Plasma ◽

Next Generation Sequencing Ngs ◽

Taqman Qpcr Assay ◽

Taqman Qpcr

Plasma cell-free DNA (cfDNA) is frequently analyzed using liquid biopsy to investigate cancer markers. We hypothesized that this concept might be applicable in exercise physiology. Here, we aimed to identify specific cfDNA (spcfDNA) sequences in the plasma of healthy humans using next-generation sequencing (NGS) and clearly define the dynamics regarding spcfDNA-fragment levels upon extreme exercises, such as running a full marathon. NGS analysis was performed using cfDNA of pooled plasma collected from healthy participants. We confirmed that the TaqMan-qPCR assay had high sensitivity and found that the spcfDNA sequence abundance was 16,600-fold higher than that in a normal genomic region. We then used the TaqMan-qPCR assay to investigate the dynamics of spcfDNA-fragment levels upon running a full marathon. The spcfDNA fragment levels were significantly increased post-marathon. Furthermore, spcfDNA fragment levels were strongly correlated with white blood cell and plasma myoglobin concentrations. These results suggest the spcfDNA fragments identified in this study were highly sensitive as markers of extreme physical stress. The findings of this study may provide new insights into exercise physiology and genome biology in humans.

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Diagnostic Tests Can Stem the Threat of Antimicrobial Resistance: Infectious Disease Professionals Can Help

Clinical Infectious Diseases ◽

10.1093/cid/ciaa1527 ◽

2020 ◽

Author(s):

Dana Trevas ◽

Angela M Caliendo ◽

Kimberly Hanson ◽

Jaclyn Levy ◽

Christine C Ginocchio

Keyword(s):

Infectious Disease ◽

Antimicrobial Resistance ◽

Diagnostic Tests ◽

Healthcare Providers ◽

Federal Funding ◽

Antibiotic Use ◽

Advisory Council ◽

Resistant Bacteria ◽

Research Development ◽

Antibiotic Resistant

Abstract Uptake of existing diagnostics to identify infections more accurately could minimize unnecessary antibiotic use and decrease the growing threat of antibiotic resistance. The Infectious Diseases Society of America (IDSA) and the Presidential Advisory Council on Combating Antibiotic-Resistant Bacteria (PACCARB) agree that, to improve uptake of existing diagnostics, healthcare providers, health systems, and payors all need better clinical and economic outcomes data to support use of diagnostic tests over empiric use of antibiotics, providers need better tools and education about diagnostic tests, and diagnostics developers need federal funding in the absence of a viable diagnostics market. Recommendations from PACCARB and the IDSA are amplified. Incentives for—and challenges to—diagnostics research, development, and uptake are summarized. Advocacy opportunities are given for infectious disease professionals to join the fight against antimicrobial resistance.

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