Abstract
Parafilaroides decorus, also known as sea lion lungworm, is a metastrongyloid nematode that infects otariid hosts, such as the charismatic California sea lion, Zalophus californianus. P. decorus causes bronchointerstitial pneumonia, respiratory distress, reduced ability to swim, dive and hunt and as a result, increased mortality particularly in young animals. Respiratory disease is a leading cause of stranding and admission to rehabilitation centers on the Pacific coast. Low-coverage genomic sequencing of four P. decorus individuals analyzed through Galaxy's RepeatExplorer identified a novel repeat DNA family we employed to design a sensitive quantitative PCR (qPCR) assay for diagnosing infections from fecal or sputum samples. The assay detects as little as 10 fg of P. decorus DNA and a linear regression model developed using a standard curve can be used to estimate the concentration of P. decorus DNA in a sample, ± 0.015 ng. This knowledge can be leveraged to estimate the level of parasite burden, which can be used to design improved treatments for animals in rehabilitation. Improved treatment of infections will aid in more animals being successfully released back into the wild.
Highlights
To the detriment of much marine life, global warming is predicted to increase the range, prevalence, and virulence of aquatic parasites (Harvell et al, 2002)
Our goal is to create sensitive, species-specific assays for diagnosis of pre-patent infection in California sea lion (Zalophus californianus) patients who present with respiratory disease and to assist clinicians in monitoring the efficacy of anthelminthic treatment (Field et al, 2018)
Parafilaroides decorus and other nematode parasites were obtained from stranded, deceased otariids, while fecal and sputum samples were obtained from rehabilitating otariids collected by The Marine Mammal Center (TMMC, Sausalito, CA; Table 1)
Summary
To the detriment of much marine life, global warming is predicted to increase the range, prevalence, and virulence of aquatic parasites (Harvell et al, 2002) Accurate identification of these parasites can allow for better assessment and treatment of pathogenic threats to wild marine mammals and treat infections of those being cared for in rehabilitation facilities. By using a repeatbased quantitative PCR (qPCR) assay with a higher number of genome targets, assays are more sensitive but allow for the estimation of parasite burden (Pilotte et al, 2019). In nematodes, these non-coding repeats evolve quickly enough that they tend to be species-specific (Pilotte et al, 2016). The California sea lion (Zalophus californianus), has been rated
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