EARLY VIEW
METHOD DEVELOPMENT FOR RAPID STREAMSIDE
ASSESSMENT OF FRESHWATER MUSSEL (ORDER
UNIONIDA) HEMOCYTE CONCENTRATION AND
MORPHOMETRICS
Madison E. Mueller, W. Gregory Cope, Christopher B. Eads,
Loretta M. Lutackas, and Dorian J. Hayes
ABSTRACT
Contemporary freshwater mussel population declines often lack discriminant explanations, and
diagnoses are commonly post hoc, subjective, expensive, and/or lethal. To avoid further compromising
imperiled populations, we need sublethal metrics and biomarkers that identify early warning signs of
infection, stress responses, and deviations from physiological homeostasis or potential resilience to
environmental pressures. Immune modulators can be differentiated by quantifying multiple
physiological endpoints based on hemocyte form and function. Spontaneous hemocyte aggregation, cell
instability, travel times from mussel collection sites, and a lack of access to costly instrumentation
currently limit immune status assessments. Widespread capacity to understand immune responses to
disease, toxicants, and other stressors relies on effective hemolymph handling and preservation
techniques. In this study we optimized methods to measure total hemocyte concentration and stability
across five antiaggregants (Alsever’s Solution, K2EDTA and sodium citrate Microtainers, Isoton II, and
Leibovitz’s L15) over time using a Beckman Coulter Multisizer 4e (a laboratory benchtop unit) to
validate results from aMillipore Scepter (a portable, field-ready hand-held unit). K2EDTA Microtainers
held on ice for up to 6 h preserved hemolymph most effectively based on consistency in total hemocyte
concentration and morphometrics over time. There was a highly significant agreement (R2 ¼ 0.9156, P ,
0.0001) in total hemocyte concentration between the Multisizer 4e and Millipore Scepter. Portable and
compact cell counters provide a less expensive, advantageous alternative to benchtop instruments for
instantaneous evaluation at mussel collection sites when timely quantification would otherwise require
extensive logistical planning. Our results lay the foundation for accessible streamside assessment, which
can expedite the establishment of reference ranges among species and populations and identify how
anticipated stressors such as infection and starvation alter the immune response. This approach also may
be useful for quantifying freshwater mussel health among propagation facilities and in stable or declining
ecosystems.
GENETIC VARIATION IN AN INVASIVE CLAM (FORM B:
CORBICULA LARGILLIERTII) ACROSS ILLINOIS, USA
Brenden Mattfolk, Julia W. Bondi Nicholas J. Iacaruso, Jeremy Tiemann,
Mark A. Davis, and Joel B. Corush
ABSTRACT
Population genetics is an important tool for understanding the source, establishment, and subsequent
spread of invasive species. It is particularly pertinent for invasive species that go unrecognized or are
understudied due to their limited detection, cryptic diversity, and/or overall lack of interest. One genus of
invasive clams, Corbicula, has been a particularly successful invader, both globally and in the USA, but has
received less attention than other aquatic invaders (e.g., invasive carp or dreissenid mussels). Herein, we
address the genetic diversity of one of the three invasive Corbicula species—Form B: Corbicula largillierti—
by using the 28S nuclear and cytochrome c oxidase subunit 1 (COI) mitochondrial gene in 74 individuals
from six localities across multiple watersheds in Illinois, USA.We identified spatial structure in both genes
and revealed an overall high level of genetic diversity and differentiation among localities. This information
can be useful for addressing questions pertaining to propagule pressure, local spread, and overall genetic
variation of this otherwise understudied invasive species.
CHARACTERIZATION OF THE NATIVE FRESHWATER
MUSSEL ASSEMBLAGE ACROSS A REACH OF THE
RAPPAHANNOCK RIVER, VIRGINIA
Traci P. DuBose1, Jason Rohweder, Catherine Gatenby, Brian Watson,
and Teresa J. Newton
ABSTRACT
Native freshwater mussels occur in rivers of all sizes, yet much of the literature focuses on small- to
medium-sized rivers, likely because of logistic challenges of sampling deep, fast, and turbid large rivers. As
large-river surveys become more common, the information resulting from a systematic sampling design can
provide baseline data on population sizes to inform decision making by resource managers. We collected
baseline data onmussel resources in the Rappahannock River, Virginia, in August 2024. This Atlantic coast
river, which drains into the Chesapeake Bay, is ~320 km long, up to 1.6 km wide, and up to 35 m deep.We
conducted a systematic quantitative survey across the 75-km reach of the river from Fredericksburg to
Leedstown, Virginia. Divers obtained 187 whole substrate samples to estimate the species richness, density,
population size, size distribution, community composition, and longitudinal spatial distribution of the native
mussel assemblage. Six species were found, and the community was dominated by Eastern Elliptio (Elliptio
complanata).Mean reach density (6 1 standard error) and population sizewas 1.25 (0.21)musselsm22 and
19.3 (3.3) million mussels, respectively. The presence of few fresh dead mussels indicated low to moderate
natural mortality. Shell length ranged from 32 to 122 mm, and multiple size classes were observed
in E. complanata. Species richness and density had similar longitudinal patterns, with the highest
values in themiddle of the sampled reach (river kilometers 60–95). These data provide resource managers
with baseline information on the spatial distribution, density, length demography, diversity, and evenness
of native freshwater mussels across this reach of the Rappahannock River. Identi!cation of mussel
occurrence, density, and evidence of reproduction can be used by managers to aid conservation and
management decisions such as prioritizing areas for reintroductions and habitat restoration.
UPPER THERMAL TOLERANCE OF THE WESTERN
RIDGED MUSSEL (GONIDEA ANGULATA), WESTERN
PEARLSHELL (MARGARITIFERA FALCATA), AND
WESTERN SCULPIN SPECIES (COTTUS SPP.)
Rachael Valeria1, Alexa Maine, Christine Parent, and James J. Nagler
ABSTRACT
Freshwater mussels are ecologically and culturally important and globally have the highest extinction
and imperilment rates of any animal. The Western Ridged Mussel (Gonidea angulata) and the Western
Pearlshell (Margaritifera falcata) are experiencing die-offs across their native ranges, and increasing
water temperature is a possible contributing factor. To host their larval life stage, G. angulata uses the
gills of sculpins (Cottus spp.) and M. falcata uses the gills of salmonids (Oncorhynchus spp.). The purpose
of this laboratory study was to determine the upper thermal tolerances of G. angulata and M. falcata and
two sculpin species, Margined Sculpin (Cottus marginatus) and Paiute Sculpin (Cottus beldingi), by using
sublethal acute and chronic survival tests. The acute test consisted of increasing the water temperature
until critical thermal maximum (CTmax) temperature behaviors were observed. The mean CTmax
temperature was 34.1°C for G. angulata and 33.2°C for M. falcata, and the mean for the two Cottus spp.
was 28.9°C. Based on the acute test results, G. angulata and M. falcata have a higher CTmax temperature
than their host fish. The chronic test gradually increased the water temperature surrounding the test
organism to 2°C below the CTmax temperature for each species. This temperature was maintained for
7 d, during which survival was recorded. All mussels perished, whereas 58.3% of the sculpins survived
the chronic test. Survival curves from the chronic test showed that G. angulata survived longer and are
more tolerant to prolonged elevated temperatures than M. falcata. These thermal tolerance tests
demonstrate that a thermal separation between mussel and host fish is possible, thus limiting mussel
reproduction as climate change leads to warming surface waters. In addition, tolerance to prolonged
elevated temperature varies between these western mussel species.
