Ghost Factors of Laboratory Carbonate Chemistry Are Haunting Our Experiments.

AbstractFor many historical and contemporary experimental studies in marine biology, seawater carbonate chemistry remains a ghost factor, an uncontrolled, unmeasured, and often dynamic variable affecting experimental organisms or the treatments to which investigators subject them. We highlight how environmental variability, such as seasonal upwelling and biological respiration, drive variation in seawater carbonate chemistry that can influence laboratory experiments in unintended ways and introduce a signal consistent with ocean acidification. As the impacts of carbonate chemistry on biochemical pathways that underlie growth, development, reproduction, and behavior become better understood, the hidden effects of this previously overlooked variable need to be acknowledged. Here we bring this emerging challenge to the attention of the wider community of experimental biologists who rely on access to organisms and water from marine and estuarine laboratories and who may benefit from explicit considerations of a growing literature on the pervasive effects of aquatic carbonate chemistry changes.

Eelgrass pathogen Labyrinthula zosterae synthesizes essential fatty acids.

Negative consequences of parasites and disease on hosts are usually better understood than their multifaceted ecosystem effects. The pathogen Labyrinthula zosterae (Lz) causes eelgrass wasting disease but has relatives that produce large quantities of nutritionally valuable long-chain polyunsaturated fatty acids (LCPUFA) such as docosahexaenoic acid (DHA). Here we quantify the fatty acids (FA) of Lz cultured on artificial media, eelgrass-based media, and eelgrass segments to investigate whether Lz may similarly produce LCPUFA. We also assess whether field-collected lesions show similar FA patterns to laboratory-inoculated eelgrass. We find that Lz produces DHA as its dominant FA along with other essential FA on both artificial and eelgrass-based media. DHA content was greater in both laboratory-inoculated and field-collected diseased eelgrass relative to their respective controls. If Lz's production scales in situ, it may present an unrecognized source of LCPUFA in eelgrass ecosystems.

Environment, dosage, and pathogen isolate moderate virulence in eelgrass wasting disease.

Eelgrass wasting disease, caused by the marine pathogen Labyrinthula zosterae, has the potential to devastate important eelgrass habitats worldwide. Although this host-pathogen interaction may increase under certain environmental conditions, little is known about how disease severity is impacted by multiple components of a changing environment. In this study, we investigated the effects of variation in 3 different L. zosterae isolates, pathogen dosage, temperature, and light on severity of infections. Severity of lesions on eelgrass varied among the 3 different isolates inoculated in laboratory trials. Our methods to control dosage of inoculum showed that disease severity increased with pathogen dosage from 104 to 106 cells ml-1. In a dosage-controlled light and temperature 2-way factorial experiment consisting of 2 light regimes (diel light cycle and complete darkness) and 2 temperatures (11 and 18°C), L. zosterae cell growth rate in vitro was higher at the warmer temperature. In a companion experiment that tested the effects of light and temperature in in vivo inoculations, disease severity was higher in dark treatments and temperature was marginally significant. We suggest that the much greater impact of light in the in vivo inoculation experiment indicates an important role for plant physiology and the need for photosynthesis in slowing severity of infections. Our work with controlled inoculation of distinct L. zosterae isolates shows that pathogen isolate, increasing dosage of inoculum, increasing temperature, and diminishing light increase disease severity, suggesting L. zosterae will cause increased damage to eelgrass beds with changing environmental conditions.