M. Christopher Barnhart

Adaptations to host infection and larval parasitism in Unionoida

Abstract Freshwater mussel larval parasitism of fish is unique among bivalves. The relationship is primarily phoretic rather than nutritive; only the smallest glochidia and the haustorial larva grow substantially while on the host. Growth of the smallest larvae suggests a lower functional size limit of ∼150 μm for the juvenile stage. Most Ambleminae, the most diverse North American clade, infect host gills by attracting feeding fish. Many species of Pleurobemini and some Lampsilini release conglutinates of eggs and larvae that resemble host food items. Many Lampsilini and a few Quadrulini use mantle modifications to attract host fish to the female. The mantle of some Quadrulini forms a posterior chamber that holds glochidia for immediate release in response to host fish. In many Lampsilini, mantle flap lures and a protrusible marsupium promote attack by the host fish and direct extraction of glochidia from the marsupium by the host. Host extraction of glochidia from the brooding female might have favored the evolution of long-term brooding in Lampsilini because glochidia need not be released by the female to encounter the host. A remarkable derivative of the host extraction strategy evolved in Epioblasma, which catch fish between the valves and release glochidia directly to the trapped host before releasing it. Host specificity is a critical feature of the evolutionary diversification and conservation biology of Unionoida. As temporary parasites, mussels must primarily evade the innate immune responses of the host, rather than the adaptive (acquired) responses. Evolution of host specificity is associated with selective encounter of host taxa, either because of host attraction strategies or because of dominance of particular host species in the habitat. The intricate relationships between mussels and fish are easily disrupted and, thus, contribute to the imperilment of many mussel species, yet they also fascinate us and compel conservation efforts.

Differential exposure, duration, and sensitivity of unionoidean bivalve life stages to environmental contaminants

Abstract Freshwater mussels (superfamily Unionoidea) are in serious global decline and in urgent need of protection and conservation. The declines have been attributed to a wide array of human activities resulting in pollution and water-quality degradation, and habitat destruction and alteration. Linkages among poor water quality, pollutant sources, and mussel decline in rivers and streams have been associated with results of laboratory-based tests of specific pollutants. However, uncertainties remain about the relationship of laboratory data to actual contaminant exposure routes for various mussel species, life stages, and in the habitats occupied during these exposures. We evaluated the pathways of exposure to environmental pollutants for all 4 life stages (free glochidia, encysted glochidia, juveniles, adults) of unionoidean mussels and found that each life stage has both common and unique characteristics that contribute to observed differences in exposure and sensitivity. Free glochidia typically are exposed only briefly (e.g., seconds to days) through surface water, whereas adults sustain exposure over years to decades through surface water, pore water, sediment, and diet. Juveniles live largely burrowed in the sediment for the first 0 to 4 y of life. Thus, sediment, pore water, and diet are the predominant exposure routes for this life stage, but surface water also might contribute to exposure during certain periods and environmental conditions. The obligate parasitic stage (encysted glochidia stage) on a host fish might be exposed from surface water while partially encysted or from toxicants in host-fish tissue while fully encysted. Laboratory methods for testing for acute and chronic exposures in water have advanced, and toxicant-specific information has increased in recent years. However, additional research is needed to understand interactions of life history, habitat, and long-term exposure to contaminants through water, pore water, sediment, and diet so that the risks of environmental exposures can be properly assessed and managed.

Acute and chronic toxicity of technical-grade pesticides to glochidia and juveniles of freshwater mussels (unionidae)

Chemical contaminants are among many potential factors involved in the decline of freshwater mussel populations in North America, and the effects of pesticides on early life stages of unionid mussels are largely unknown. The objective of this study was to determine the toxicity of technical-grade current-use pesticides to glochidia and juvenile life stages of freshwater mussels. We performed acute toxicity tests with glochidia (five species) and juveniles (two species) exposed to a suite of current-use pesticides including herbicides (atrazine and pendimethalin), insecticides (fipronil and permethrin), and a reference toxicant (NaCl). Because of limited availability of test organisms, not all species were tested with all pesticides. Toxicity tests with fungicides (chlorothalonil, propiconazole, and pyraclostrobin) were performed with one species (Lampsilis siliquoidea). Lampsilis siliquoidea glochidia and juveniles were highly sensitive to the fungicides tested but the technical-grade herbicides and insecticides, at concentrations approaching water solubility, were not acutely toxic to this or the other unionid species. In a 21-d chronic test with four-month-old juvenile L. siliquoidea, the 21-d median effective concentration (EC50) with atrazine was 4.3 mg/L and in atrazine treatments >or=3.8 mg/L mussel growth was significantly less than controls. The relatively high sensitivity of L. siliquoidea to chlorothalonil, propiconazole, and pyraclostrobin is similar to that reported for other aquatic organisms commonly used for toxicity testing. The relative risk associated with acute exposure of early life stages of mussels to technical-grade atrazine, pendimethalin, fipronil, and permethrin is likely low; however, survival and growth results with juvenile L. siliquoidea indicate that chronic exposure to high concentrations (>/=3.8 mg/L) of atrazine may have the potential to impact mussel populations and warrants further investigation.

Respiration and heat production by the inflorescence of Philodendron selloum Koch

During a 2-d sequence of anthesis, the spadices of the thermogenic arum lily, Philodendron selloum, regulated maximum temperature within a small range (37–44°C) by reversible thermal inhibition of respiratory heat production. This response protects the inflorescence and the attracted insects from thermal damage. Heat production by whole spadices, measured by O2 respirometry, equalled heat loss, measured by gradient layer calorimetry, which confirmed the heat equivalence of O2 consumption (20.4 J ml-1). This also indicated that there was no net phosphorylation during thermogenesis, heat production being the primary function of high rates of respiration. The sterile male florets consumed about 30 ml g-1 h-1 and the average 124-g spadix produced about 7 W to maintain a 30°C difference between spadix and ambient temperature. Most of the energy for thermogenesis is present in the florets before anthesis. Despite high respiratory rates, thermogenesis is an energetically inexpensive component of the reproductive process.

Cross-resistance of largemouth bass to glochidia of unionid mussels

We tested whether host fish that acquired resistance to glochidia of one mussel species were cross-resistant to glochidia of other species. Largemouth bass (Micropterus salmoides) were primed with 4–5 successive infections of glochidia of Lampsilis reeveiana. The percentage of attached glochidia that survived and transformed to the juvenile stage (transformation success) was compared between primed fish and naïve controls. Transformation success of L. reeveiana, Lampsilis abrupta, Villosa iris, and Utterbackia imbecillis was significantly lower on primed fish (37.8%, 43.5%, 67.0%, and 13.2%, respectively) than on control fish (89.0%, 89.7%, 90.0%, and 22.2% respectively). Immunoblotting was used to analyze the binding of serum antibodies from primed fish with glochidia proteins. Antibodies bound to glochidia proteins of similar molecular weight from L. reeveiana and L. abrupta. Bound proteins of V. iris differed in molecular weight from those of the Lampsilis species. There was no binding to specific glochidia proteins of U. imbecillis or Strophitus undulatus. Our results indicate that host-acquired resistance can extend across mussel genera and subfamilies and might involve both specific and nonspecific mechanisms. Understanding the specificity of acquired resistance of hosts to glochidia could enhance understanding of the evolutionary and ecological relationships between mussels and their host fishes.

Upper thermal tolerances of early life stages of freshwater mussels

Abstract Freshwater mussels (order Unioniformes) fulfill an essential role in benthic aquatic communities, but also are among the most sensitive and rapidly declining faunal groups in North America. Rising water temperatures, caused by global climate change, industrial discharges, drought, or land development, could further challenge imperiled unionid communities. The aim of our study was to determine the upper thermal tolerances of the larval (glochidia) and juvenile life stages of freshwater mussels. Glochidia of 8 species of mussels were tested: Lampsilis siliquoidea, Potamilus alatus, Ligumia recta, Ellipsaria lineolata, Lasmigona complanata, Megalonaias nervosa, Alasmidonta varicosa, and Villosa delumbis. Seven of these species also were tested as juveniles. Survival trends were monitored while mussels held at 3 acclimation temperatures (17, 22, and 27°C) were exposed to a range of common and extreme water temperatures (20–42°C) in standard acute laboratory tests. The average median lethal temperature (LT50) among species in 24-h tests with glochidia was 31.6°C and ranged from 21.4 to 42.7°C. The mean LT50 in 96-h juvenile tests was 34.7°C and ranged from 32.5 to 38.8°C. Based on comparisons of LT50s, thermal tolerances differed among species for glochidia, but not for juveniles. Acclimation temperature did not affect thermal tolerance for either life stage. Our results indicate that freshwater mussels already might be living close to their upper thermal tolerances in some systems and, thus, might be at risk from rising environmental temperatures.

Survival of Immersion and Anoxia by Larval Tiger Beetles, Cicindela togata

Abstract The sedentary terrestrial larvae of the tiger beetle, Cicindela togata, inhabit areas that are often flooded for days or weeks. We tested the ability of these larvae to survive immersion and anoxia. Maximum survival time of immersed, anoxic C. togata was 6 days at 25 C. Time to 50% mortality (LT50) in these conditions was 85.9 ± 23.5 h. Survival times were more than eight times longer than those of similarly treated larvae of Tenebrio molitor (LT50 10.1 ± 3.2 h). Similar or somewhat longer survival times were observed in larvae of C. togata exposed to an anoxic nitrogen atmosphere (LT50 102 ± 31 h) and of T. molitor (LT5014.4 ± 6.5 h). At 10 C, LT50 of C. togata in anoxic atmospheres exceeded 10 days. Tiger beetle larvae are physiologically capable of surviving several days of inundation during floods without mechanisms to prevent burrow flooding. By entering a quiescent state, C. togata larvae survive much longer periods of anoxia than has been previously reported for terrestrial insect larvae.

Control of acid-base status in active and dormant land snails, Otala lactea (Pulmonata, Helicidae)

SummaryControl of extracellular acid-base status was examined during activity and dormancy inOtala lactea (Pulmonata, Helicidae). Active snails showed little variation in hemolymph pH and

Effects of Temperature, pH, and CO 2 on Transformation of the Glochidia of Anodonta suborbiculata on Fish Hosts and in Vitro

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