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Andrew P. Kinziger

Publications and source records attributed to Andrew P. Kinziger.

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Quantitative PCR detection of endangered diamond darter Crystallaria Cincotta in environmental DNA: Employing locked nucleic acids and blocking probe for specificity

This study presents a quantitative PCR (qPCR) assay for the detection of the endangered diamond darter Crystallaria cincotta from environmental DNA (eDNA) in water samples. The assay design is based on an alignment of mitochondrial cytochrome b DNA sequences from 58 individuals representing 25 percid species. Leveraging genetic differences, a species-specific qPCR assay was designed, incorporating alocked nucleic acid (LNA)-enriched probe and a secondary blocking probe to enhance specificity. The assay targets a 93-base pair fragment that includes a diagnostic single nucleotide polymorphism in the probe region; combined with multiple primer mismatches, this provides specificity for distinguishing C. cincotta from other sympatric percid species. Specificity was validated by testing genomic DNA from 16 percid species and synthetic templates, confirming no cross-reactivity. Performance metrics, including the standard curve, qPCR efficiency, limit of detection, and limit of quantification, are reported. The qPCR assay exhibited sufficient sensitivity to detect C. cincotta eDNA in environmental water samples collected from occupied riverine habitats. This study illustrates the effectiveness of LNA-enriched and blocking probes in developing species-specific qPCR assays for eDNA applications, demonstrating their utility in accurately distinguishing closely related species within diverse fish communities.

Conservation Genetics Resources

Testing and extension of a sea lamprey feeding model

A previous model of feeding by sea lamprey Petromyzon marinus predicted energy intake and growth by lampreys as a function of lamprey size, host size, and duration of feeding attachments, but it was applicable only to lampreys feeding at 10°C and it was tested against only a single small data set of limited scope. We extended the model to other temperatures and tested it against an extensive data set (more than 700 feeding bouts) accumulated during experiments with captive sea lampreys. Model predictions of instantaneous growth were highly correlated with observed growth, and a partitioning of mean squared error between model predictions and observed results showed that 88.5% of the variance was due to random variation rather than to systematic errors. However, deviations between observed and predicted values varied substantially, especially for short feeding bouts. Predicted and observed growth trajectories of individual lampreys during multiple feeding bouts during the summer tended to correspond closely, but predicted growth was generally much higher than observed growth late in the year. This suggests the possibility that large overwintering lampreys reduce their feeding rates while attached to hosts. Seasonal or size-related shifts in the fate of consumed energy may provide an alternative explanation. The lamprey feeding model offers great flexibility in assessing growth of captive lampreys within various experimental protocols (e.g., different host species or thermal regimes) because it controls for individual differences in feeding history.

Transactions of the American Fisheries Society