Donovan P. German

Digestive Enzyme Activities in Herbivorous and Carnivorous Prickleback Fishes (Teleostei: Stichaeidae): Ontogenetic, Dietary, and Phylogenetic Effects

We measured the activities of eight digestive enzymes in four species of herbivorous and carnivorous prickleback fishes and determined the effects of ontogeny, diet, and phylogeny on these enzyme activities. Of the four species, Cebidichthys violaceus and Xiphister mucosus shift to a more herbivorous diet as they grow (≥45 mm SL [standard length]), whereas Xiphister atropurpureus and Anoplarchus purpurescens remain carnivores throughout life. Digestive enzyme activities of small (30–40 mm SL) carnivorous juveniles were compared with those of larger (60–75 mm SL) wild‐caught juveniles that had consumed a natural diet and larger (60–75 mm SL) juveniles raised on a high‐protein animal diet. Cebidichthys violaceus and both species of Xiphister showed ontogenetic changes in digestive enzyme activities, whereas A. purpurescens did not. Despite dietary differences between X. atropurpureus and X. mucosus, these sister taxa displayed the most similar digestive enzyme activities from ontogenetic and dietary perspectives (high α‐amylase and lipase and low trypsin and aminopeptidase activities), and both were more similar to C. violaceus, a member of the same largely herbivorous clade, than either was to A. purpurescens, a member of an adjacent, carnivorous clade. The results support the hypothesis that phylogeny influences digestive enzyme activities in these fishes. Anoplarchus purpurescens, a carnivore with a diverse diet, showed great plasticity in enzyme activity, especially trypsin and aminopeptidase, which were elevated in this species to the highest level among the four species after consuming the high‐protein diet. These results support the hypothesis that fishes with relatively broad diets can modulate digestive enzyme activities in response to changes in dietary composition.

Substrate concentration and enzyme allocation can affect rates of microbial decomposition.

A large proportion of the worlds carbon is stored as soil organic matter (SOM). However, the mechanisms regulating the stability of this SOM remain unclear. Recent work suggests that SOM may be stabilized by mechanisms other than chemical recalcitrance. Here, we show that the mineralization rate of starch, a plant polymer commonly found in litter and soil, is concentration dependent, such that its decomposition rate can be reduced by as much as 50% when composing less than approximately 10% of SOM. This pattern is largely driven by low activities of starch-degrading enzymes and low inducibility of enzyme production by microbial decomposers. The same pattern was not observed for cellulose and hemicellulose degradation, possibly because the enzymes targeting these substrates are expressed at constitutively high levels. Nevertheless, given the heterogeneous distribution of SOM constituents, our results suggest a novel low-concentration constraint on SOM decomposition that is independent of chemical recalcitrance. These results may help explain the stability of at least some SOM constituents, especially those that naturally exist in relatively low concentrations in the soil environment.

Evolution of herbivory in a carnivorous clade of minnows (teleostei: cyprinidae): effects on gut size and digestive physiology.

We constructed a phylogeny for 10 minnow species (family Cyprinidae) previously revealed to be members of sister genera with different dietary affinities and used the phylogeny to examine whether the evolution of digestive tract size and physiology is correlated with the evolution of diet in these fishes. We studied a total of 11 taxa: four herbivorous species in the genus Campostoma and six largely carnivorous species in the genus Nocomis, including two populations of Nocomis leptocephalus, the carnivorous Chattahoochee River drainage population and the herbivorous Altamaha River drainage population. Thus, we were able to compare digestive tract size and physiology among sister genera (Campostoma and Nocomis) and among sister taxa (N. leptocephalus Chattahoochee and N. leptocephalus Altamaha) in dietary and phylogenetic contexts. The herbivorous taxa had longer digestive tracts and higher activity of the carbohydrases amylase and laminarinase in their guts, whereas the carnivorous species had higher chitinase activity. Phylogenetic independent‐contrast analysis suggested that the evolution of amylase and chitinase activities was correlated with the evolution of diet in these species, whereas trypsin and lipase activities showed no pattern associated with diet or phylogenetic history. Concentrations of short‐chain fatty acids were low in all taxa, indicating that these fishes rely largely on endogenous digestive mechanisms to subsist on their respective diets. Subtle differences in tooth shape were observed between species in the two genera. Overall, our results suggest that dietary specialization can be observed on the level of anatomy and physiology of the digestive tracts of fishes but that such differences are most appropriately viewed in comparisons of closely related species with different diets.

Feast to famine: The effects of food quality and quantity on the gut structure and function of a detritivorous catfish (Teleostei: Loricariidae)

The gastrointestinal (GI) tract and associated organs are some of the most metabolically active tissues in an animal. Hence, when facing food shortages or poor food quality, an animal may reduce the size and function of their GI tract to conserve energy. We investigated the effects of prolonged starvation and varying food quality on the structure and function of the GI tract in a detritivorous catfish, Pterygoplichthys disjunctivus, native to the Amazonian basin, which experiences seasonal variation in food availability. After 150 days of starvation or consumption of a wood-diet too low in quality to meet their energetic needs, the fish reduced the surface area of their intestines by 70 and 78%, respectively, and reduced the microvilli surface area by 52 and 27%, respectively, in comparison to wild-caught fish consuming their natural diet and those raised in the laboratory on a high-quality algal diet. Intake and dietary quality did not affect the patterns of digestive enzyme activity along the guts of the fish, and the fish on the low-quality diet had similar mass-specific digestive enzyme activities to wild-caught fish, but lower summed activity when considering the mass of the gut. Overall, P. disjunctivus can endure prolonged starvation and low food quality by down-regulating the size of its GI tract.

Stable carbon and nitrogen incorporation in blood and fin tissue of the catfish Pterygoplichthys disjunctivus (Siluriformes, Loricariidae)

AbstractA feeding trial was performed in the laboratory with the catfish species Pterygoplichthys disjunctivus to determine stable carbon (13C) and nitrogen (15 N) turnover rates and discrimination factors in non-lethally sampled tissues (red blood cells, plasma solutes, and fin). A second feeding trial was conducted to determine what P. disjunctivus could assimilate from low-quality wood-detritus—refractory polysaccharides (e.g., cellulose), or soluble wood-degradation products inherent in wood-detritus. This was performed by feeding the fish an artificial wood-detritus diet with fibrous (δ13C = −26.36‰; δ15  N = 2.13‰) and soluble portions (δ13C = −11.82‰; δ15  N = 3.39‰) that had different isotopic signatures and monitoring the dynamics of isotopic incorporation in the different tissues over time. Plasma solutes turned over more quickly than red blood cells for 13C and 15 N. However, in contrast to previous studies of juvenile fishes, C and N incorporation was primarily driven by catabolic tissue turnover as opposed to growth rate. Tissue-diet discrimination factors for 15 N varied from 4.08 to 5.17‰, whereas they were <2‰ for 13C (and less than 0.3‰ for plasma and red blood cells). The results of trial two suggested that P. disjunctivus could not assimilate refractory polysaccharides. Moreover, the δ13C and δ15 N signatures of wild-caught P. disjunctivus from Florida confirmed their detrital trophic standing in Floridian aquatic ecosystems.

Phenotypic plasticity of gut structure and function during periods of inactivity in Apostichopus japonicus

Apostichopus japonicus is a common sea cucumber that undergoes seasonal inactivity phases and ceases feeding during the summer months. We used this sea cucumber species as a model in which to examine phenotypic plasticity of the digestive tract in response to food deprivation. We measured the body mass, gross gut morphology and digestive enzyme activities of A. japonicus before, during, and after the period of inactivity to examine the effects of food deprivation on the gut structure and function of this animal. Individuals were sampled semi-monthly from June to November (10 sampling intervals over 178 days) across temperature changes of more than 18 degrees C. On 5 September, which represented the peak of inactivity and lack of feeding, A. japonicus decreased its body mass, gut mass and gut length by 50%, 85%, and 70%, respectively, in comparison to values for these parameters preceding the inactive period. The activities of amylase, cellulase and lipase decreased by 77%, 98%, and 35% respectively, in comparison to mean values for these enzymes in June, whereas pepsin activity increased two-fold during the inactive phase. Alginase and trypsin activities were variable and did not change significantly across the 178-day experiment. With the exception of amylase and cellulase, all body size indices and digestive enzyme activities recovered and even surpassed the mean values preceding the inactive phase during the latter part of the experiment (October-November). Principal Component Analysis (PCA) utilizing the digestive enzyme activity and body size index data divided the physiological state of this cucumber into four phases: an active stage, prophase of inactivity, peak inactivity, and a reversion phase. These phases are all consistent with previously suggested life stages for this species, but our data provide more defined characteristics of each phase. A. japonicus clearly exhibits phenotypic plasticity (or life-cycle staging) of the digestive tract during its annual inactive period.

Do herbivorous minnows have “plug-flow reactor” guts? Evidence from digestive enzyme activities, gastrointestinal fermentation, and luminal nutrient concentrations

Few investigations have empirically analyzed fish gut function in the context of chemical reactor models. In this study, digestive enzyme activities, levels of gastrointestinal fermentation products [short chain fatty acids (SCFA)], luminal nutrient concentrations, and the mass of gut contents were measured along the digestive tract in herbivorous and carnivorous minnows to ascertain whether their guts function as “plug-flow reactors” (PFRs). Four of the species, Campostoma anomalum, C. ornatum, C. oligolepis, and C. pauciradii, are members of a monophyletic herbivorous clade, whereas the fifth species, Nocomis micropogon, is a carnivore from an adjacent carnivorous clade. In the context of a PFR model, the activities of amylase, trypsin and lipase, and the concentrations of glucose, protein, and lipid were predicted to decrease moving from the proximal to the distal intestine. I found support for this as these enzyme activities and nutrient concentrations generally decreased moving distally along the intestine of the four Campostoma species. Furthermore, gut content mass and the low SCFA concentrations did not change (increase or decrease) along the gut of any species. Combined with a previous investigation suggesting that species of Campostoma have rapid gut throughput rates, the data presented here generally support Campostoma as having guts that function as PFRs. The carnivorous N. micropogon showed some differences in the measured parameters, which were interpreted in the contexts of intake and retention time to suggest that PFR function breaks down in this carnivorous species.

Why can't young fish eat plants? Neither digestive enzymes nor gut development preclude herbivory in the young of a stomachless marine herbivorous fish

Most young fishes lack the ability to function as herbivores, which has been attributed to two aspects of the digestive system: elevated nitrogen demand and a critical gut capacity. We compared the digestive morphology and biochemistry of two size classes of the marine herbivore Hyporhamphus regularis ardelio, pre-ontogenetic trophic shift (pre-OTS, <100mm) and post-ontogenetic trophic shift (post-OTS, >100mm), to determine what limits the onset of herbivory and how their digestive processes fit with current models of digestion. Two gut-somatic indices comparing gut length to body length (relative gut length) and body mass (Zihlers Index) demonstrated a significant decrease (RGL 0.59→0.49, P<0.01; ZI 3.24→2.44, P<0.01) in gut length relative to body size. There was little difference in enzyme activity between the two classes, with juveniles showing similar levels of carbohydrase and lipase and less protease compared with adults, indicating that juveniles did not preferentially target nitrogen and were as capable of digesting an herbivorous diet. These findings suggest that herbivory in this fish is not limited by the function of the post-oesophageal digestive tract, but rather the ability of the pharyngeal mill to mechanically process plants. Our findings offer partial support for the current model of stomachless digestion, indicating that further refinement may be necessary.

More than one way to be an herbivore: convergent evolution of herbivory using different digestive strategies in prickleback fishes (Stichaeidae)

In fishes, the evolution of herbivory has occured within a spectrum of digestive strategies, with two extremes on opposite ends: (i) a rate-maximization strategy characterized by high intake, rapid throughput of food through the gut, and little reliance on microbial digestion or (ii) a yield-maximization strategy characterized by measured intake, slower transit of food through the gut, and more of a reliance on microbial digestion in the hindgut. One of these strategies tends to be favored within a given clade of fishes. Here, we tested the hypothesis that rate or yield digestive strategies can arise in convergently evolved herbivores within a given lineage. In the family Stichaeidae, convergent evolution of herbivory occured in Cebidichthys violaceus and Xiphister mucosus, and despite nearly identical diets, these two species have different digestive physiologies. We found that C. violaceus has more digesta in its distal intestine than other gut regions, has comparatively high concentrations (>11 mM) of short-chain fatty acids (SCFA, the endpoints of microbial fermentation) in its distal intestine, and a spike in β-glucosidase activity in this gut region, findings that, when coupled to long retention times (>20 h) of food in the guts of C. violaceus, suggest a yield-maximizing strategy in this species. X. mucosus showed none of these features and was more similar to its sister taxon, the omnivorous Xiphister atropurpureus, in terms of digestive enzyme activities, gut content partitioning, and concentrations of SCFA in their distal intestines. We also contrasted these herbivores and omnivores with other sympatric stichaeid fishes, Phytichthys chirus (omnivore) and Anoplarchus purpurescens (carnivore), each of which had digestive physiologies consistent with the consumption of animal material. This study shows that rate- and yield-maximizing strategies can evolve in closely related fishes and suggests that resource partitioning can play out on the level of digestive physiology in sympatric, closely related herbivores.

Evolution of ontogenetic dietary shifts and associated gut features in prickleback fishes (Teleostei: Stichaeidae)

We tested the hypothesis that an ontogenetic dietary shift from carnivory to herbivory or omnivory, and concomitant changes in the gut facilitating digestion of algae, are synapomorphies of the tribes Xiphisterini and Esselenichthyini in the family Stichaeidae (pricklebacks). Previous investigations have revealed that two xiphisterine pricklebacks-Xiphister mucosus and Xiphister atropurpureus-become herbivorous or omnivorous, respectively, as their bodies grow larger, and that their guts show related changes in length and function. In this study we found that, with increase in size, the basal member of the Xiphisterini, Phytichthys chirus, showed an increased proportion of algae in its diet, increased activity of α-amylase and decreased activity of aminopeptidase, all of which support the synapomorphy hypothesis. Cebidichthys violaceus, a herbivore in the Esselenichthyini, shows similar ontogenetic changes in diet and digestive tract length and physiology, but these features were not observed in two derived carnivores, Dictyosoma burgeri and Dictyosoma rubrimaculatum, within the clade. These results suggest that herbivory is isolated to C. violaceus within the Esselenichthyini. Allometric relationships of gut length as a function of body size generally follow diet within the Xiphisterini and Esselenichthyini, with herbivores having the longest guts, which become disproportionately longer than body size as the fishes grow, omnivores intermediate gut lengths, and carnivores the shortest. A carnivore from an adjacent clade, Anoplarchus purpurescens, had the shortest gut, which did not change in length relative to body length as the fish grew. Overall, our results clarify the patterns of dietary evolution within the Stichaeidae and lay the foundation for more detailed studies of dietary and digestive specialization in fishes in the family.