Production and characterization of a mouse monoclonal antibody against smallmouth bass (Micropterus dolomieu) IgM.

A murine monoclonal antibody (mAb, IgG2a) was produced for the detection of smallmouth bass (Micropterus dolomieu) immunoglobulin (IgM). The antibody is specific for IgM heavy chain and was shown to also recognize the Ig heavy chain of largemouth bass (Micropterus salmoides) and bluegill (Lepomis macrochirus) using Western Blot analysis of plasma from 9 teleost taxa. When applied to the analysis of smallmouth bass total plasma IgM using ELISA, the mAb was found to be effective when used in an inhibition kinetic assay.

In vitro immune function in laboratory-reared age-0 smallmouth bass (Micropterus dolomieu) relative to diet.

Smallmouth bass (Micropterus dolomieu) are used as an indicator species in environmental monitoring and assessment studies. However, laboratory-based studies for methods development and effector assessment are limited for this species. Nutrition, a known modulator of teleost physiological responses including immune function, is a critical knowledge-gap sometimes overlooked in the design of laboratory studies. We report the results from a study evaluating a commercially available artificial pelleted diet for bass and live feed (fathead minnows). Following a six-month diet-acclimation period, age-0 smallmouth bass were assessed using morphometric measures, histologic and immune-function end points using conventional methods, miniaturized cell isolation and assay methods as well as imaging flow cytometry. Fish on the two diets did not significantly differ in length, weight, or condition factor, indicating that growth was similar in the two groups. Histologic examination revealed relatively higher levels of macrophage aggregates and accumulation of ceroid/lipofuscin in the spleen as well as hepatocyte changes in the pellet-fed group. Leukocytes from the pellet-fed group exhibited significantly elevated bactericidal activity and significantly depressed mitogen response compared to fish fed live feed. Following exposure to a known inducer of inflammatory responses, bacterial lipopolysaccharide, responses including the transition of leukocytes to an apoptotic/necrotic condition differed significantly based on diet. Histologic findings were consistent with the occurrence of diet-related oxidative stress in the pellet-fed fish. Oxidative stress can be induced by multiple factors including environmental pollutants. For a diet to be useful in laboratory-based studies, it cannot elicit response that could also be induced by experimental treatments. To do so greatly complicates the detection of experimental effects. Until an artificial diet is developed for smallmouth bass that does not produce potentially confounding conditions for laboratory-based studies, use of a live feed appears to be the best option.

Population genetic structure and comparative diversity of smallmouth bass Micropterus dolomieu: congruent patterns from two genomes.

Genetic diversity and divergence patterns of smallmouth bass Micropterus dolomieu spawning groups are analysed across its northern native range with mtDNA cytochrome b gene sequences and eight unlinked nuclear DNA microsatellite loci. Results reveal high levels of genetic variability and significant differences in allelic representation among populations (mtDNA: mean ± s.e., HD = 0·50 ± 0·06, mean ± s.e., θST = 0·41 ± 0·02 and microsatellites: mean ± s.e. HO = 0·46 ± 0·03, mean ± s.e. θST = 0·25 ± 0·01). The distributions of 28 variant mtDNA haplotypes, which differ by an average of 3·94 nucleotides (range = 1-8), denote divergent representation among geographic areas. Microsatellite data support nine primary population groups, whose high self-assignment probabilities likewise display marked divergence. Genetic patterns demonstrate: (1) high genetic diversity in both genomes, (2) significant divergence among populations, probably resulting from natal site homing and low lifetime migration, (3) support for three post-glacial refugia that variously contributed to the current northern populations, which remain evident today despite waterway connectivity and (4) a weak yet significant genetic isolation by geographic distance pattern, indicating that other processes affect the differences among populations, such as territoriality and site fidelity.

Life-stage-specific physiology defines invasion extent of a riverine fish.

Many ecologists have called for mechanism-based investigations to identify the underlying controls on species distributions. Understanding these controls can be especially useful to construct robust predictions of how a species range may change in response to climate change or the extent to which a non-native species may spread in novel environments. Here, we link spatially intensive observations with mechanistic models to illustrate how physiology determines the upstream extent of the aquatic ectotherm smallmouth bass (Micropterus dolomieu) in two headwater rivers. Our results demonstrate that as temperatures become increasingly cold across a downstream to upstream gradient, food consumption in age 0 bass becomes increasingly constrained, and as a result, these fish become growth limited. Sufficient first summer growth of age 0 bass is essential for overwinter survival because young bass must persist from energy reserves accumulated during the summer, and those reserves are determined by body size. Our field data reveal the upstream extent of adult bass reproduction corresponds to a point in the downstream/upstream gradient where cold temperatures impair growth opportunities in young bass. This pattern was repeated in both study streams and explained why bass positioned nests twice as far upstream in the warm compared to the cold stream in the same basin. Placement of spawning nests by adult bass is likely subject to strong evolutionary selection in temperate systems: if bass spawn too far upstream, their young are unlikely to grow large enough to survive the winter. Consumption and growth in older bass (age 3-4) was far less sensitive to temperature. Based on these data, we suggest that temperature-sensitive age 0 bass constrain the upstream distribution limits of bass within temperate streams. In this study, we investigated how temperature-dependent physiology changed through the life history of a species and, in doing so, identified a climate-sensitive life-history stage that likely sets the distributional limits of all other life-history stages. We anticipate the framework developed here could be employed to identify how similar stage-specific environmental sensitivity determines distribution in many other ectothermic species.

Nodularin from benthic freshwater periphyton and implications for trophic transfer.

In 2013 and 2015, the Pennsylvania Department of Environmental Protection conducted a survey of lotic habitats within the Susquehanna, Delaware, and Ohio River basins in Pennsylvania, USA, to screen for microcystins/nodularins (MCs/NODs) in algae communities and smallmouth bass (Micropterus dolomieu). Periphyton (68 from 41 sites), juvenile whole fish (153 from 19 sites) and adult fish liver (115 from 16 sites) samples were collected and screened using an Adda enzyme-linked immunosorbent assay (ELISA). Samples that were positive for MCs/NODs were further analyzed using LC-MS/MS, including 14 variants of microcystin and NOD-R and the MMPB technique. The ELISA was positive for 47% of the periphyton collections, with NOD-R confirmed (0.7-82.2 ng g-1 d.w.) in 20 samples. NOD-R was confirmed in 10 of 15 positive juvenile whole fish samples (0.8-16.7 ng g-1 w.w.) and in 2 of 8 liver samples (1.7 & 2.8 ng g-1 w.w.). The MMPB method resulted in total MCs/NODs measured in periphyton (2.2-1269 ng g-1 d.w.), juvenile whole fish (5.0-210 ng g-1 d.w.) and adult livers (8.5-29.5 ng g-1 d.w.). This work illustrates that NOD-R is present in freshwater benthic algae in the USA, which has broader implications for monitoring and trophic transfer.