M.C. van Riel

Trophic relationships in the Rhine food web during invasion and after establishment of the Ponto-Caspian invader Dikerogammarus villosus

The Rhine ecosystem is highly influenced by anthropogenic stresses from pollution, intensive shipping and increased connectivity with other large European rivers. Canalization of the Rhine resulted in a reduction of heterogeneity to two main biotopes: sandy streambeds and riverbanks consisting of groyne stones. Both biotopes are heavily subjected to biological invasions, affecting the rivers food web structure. The Ponto-Caspian amphipods, Chelicorophium curvispinum and Dikerogammarus villosus, have exerted the highest impact on this food web. The filterfeeding C. curvispinum dominated the Rhine food web on the stones in 1998, swamping the stone substrata with mud. However, in 2001 it decreased in numbers, most likely due to top-down regulation caused by increased parasitic and predatory pressure of other more recently invaded Ponto-Caspian species. D. villosus showed a fast population increase after its invasion and particularly influenced the macroinvertebrate community on the stones by predaceous omnivory. This species seemed to have maintained its predatory level after its population established. Effects of these mass invaders on the macroinvertebrate community of sandy streambeds in the Rhine are unclear. Here, low densities of macroinvertebrates were observed with the Asiatic clam, Corbicula fluminea, as most abundant species. Stable isotope values of food webs from the stones and sand in 2001 were similar. Aquatic macrophytes are nearly absent and the food web is fuelled by phytoplankton and particulate organic matter, originating from riparian vegetation as indicated by similar δ13C values. Omnivores, filter-, deposit-, and detritus-feeders are the primary and secondary macroinvertebrate consumers and function as keystone species in transferring energy to higher trophic levels. Invaders comprise 90% of the macroinvertebrate numbers, and can be considered ecosystem engineers determining the functional diversity and food web structure of the Rhine by either bottom-up or top-down regulation.

Exotic and indigenous freshwater gammarid species: physiological tolerance to water temperature in relation to ionic content of the water

Bioinvasions by closely related species often lead to niche competition between exotic and indigenous species. The outcome of this competition is partly determined by differences in physiological tolerance of the competing species to the environmental conditions of the colonised habitat. Physiological tolerance of the invading gammarid species Gammarus tigrinus, Echinogammarus ischnus and Dikerogammarus villosus and the indigenous gammarid species Gammarus pulex, Gammarus roeseli and Gammarus fossarum from Dutch waters was studied in the laboratory by comparing their pleopod beats at rest at different water temperatures, which reflect the gammarids oxygen consumption. Pleopod beat frequencies increased from a minimum ventilatory activity of 0 beats per minute at 1 °C to maximum activity of up to 300 beats per minute at temperatures between 25 °C and 35 °C. At the state of maximum activity, a further increase in temperature was followed by a strong decrease in pleopod beat frequency, indicating acute stress, and subsequently mortality. Frequency response patterns of invading and indigenous gammarids were found to be highly similar, indicating a wide tolerance to temperature for all species. The tolerance of D. villosus, however, was reduced in brook water, indicating a lower competitive ability in relatively ion-poor water. G. tigrinus survived at higher temperatures in the more ion-rich, polluted waters than the indigenous gammarids, indicating a wider physiological tolerance and thus a higher competitive ability in these waters.

Invader-invader interactions in relation to environmental heterogeneity leads to zonation of two invasive amphipods, Dikerogammarus villosus (Sowinsky) and Gammarus tigrinus Sexton: Amphipod pilot species project (AMPIS) report 6

As biological invasions continue, interactions occur not only between invaders and natives, but increasingly new invaders come into contact with previous invaders. Whilst this can lead to species replacements, co-existence may occur, but we lack knowledge of processes driving such patterns. Since environmental heterogeneity can determine species richness and co-existence, the present study examines habitat use and its mediation of the predatory interaction between invasive aquatic amphipods, the Ponto-Caspian Dikerogammarus villosus and the N. American Gammarus tigrinus. In the Dutch Lake IJsselmeer, we found broad segregation of D. villosus and G. tigrinus by habitat type, the former predominating in the boulder zone and the latter in the soft sediment. However, the two species co-exist in the boulder zone, both on the short and longer terms. We used an experimental simulation of habitat heterogeneity and show that both species utilize crevices, different sized holes in a plastic grid, non-randomly. These amphipods appear to optimise the use of holes with respect to their ‘C-shape’ body size. When placed together, D. villosus adults preyed on G. tigrinus adults and juveniles, while G. tigrinus adults preyed on D. villosus juveniles. Juveniles were also predators and both species were cannibalistic. However, the impact on G. tigrinus of the superior intraguild predator, D. villosus, was significantly reduced where experimental grids were present as compared to absent. This mitigation of intraguild predation between the two species in complex habitats may explain the co-existence of these two invasive species.

Immunocytochemistry and in situ hybridization of neuropeptide Y in the hypothalamus of Xenopus laevis in relation to background adaptation

The amphibian Xenopus laevis is able to adapt to a dark background by releasing melanophore-stimulating hormone from the pars intermedia of the pituitary gland. The inhibition of melanophore-stimulating hormone release is accomplished by neuropeptide Y-containing axons innervating the pars intermedia. To determine the production site of neuropeptide Y involved in this inhibitory control, the distribution of neuropeptide Y in the brain has been investigated by immunocytochemistry and in situ hybridization. Immunoreactive cell bodies were visualized in, among others, the ventromedial and posterior thalamic nuclei, and the suprachiasmatic and ventral infundibular hypothalamic nuclei. A positive hybridization signal with a Xenopus-specific probe for preproneuropeptide Y-RNA was found in the diencephalic ventromedial thalamic nucleus and in the suprachiasmatic nucleus. With both immunocytochemistry and in situ hybridization, suprachiasmatic neurons appeared to be stained only in animals adapted to a white background; animals adapted to a black background showed no staining. Quantitative image analysis revealed that this effect of background adaptation is specific for suprachiasmatic neurons because no effect could be demonstrated of the background light condition on the ventral infundibular nucleus (immunocytochemistry) or the ventromedial thalamic nucleus (in situ hybridization). These results indicate that neurons in the suprachiasmatic nucleus enable the adaptation of X. laevis to a white background, by producing and releasing neuropeptide Y that inhibits the release of melanophore-stimulating hormone from the melanotrope cells in the pars intermedia of the pituitary gland.

Interference competition between alien invasive gammaridean species

The relative abundances of gammaridean species in the river Rhine have profoundly changed since the invasion of Dikerogammarus villosus in 1994/1995. This study tested whether these changes in gammaridean dominance could have been determined by interspecific competition and unequal mortality, for example by intraguild predation (IGP). Single and two species tests have been carried out in aquariums provided with all substrata present in the main channel of the Rhine. Changes in substratum choice, increased swimming activity and increased mortality of a species were used as indicators of interspecific competition during interaction between gammaridean species. Interspecific competition and mortality between the most abundant invasive gammaridean species in the Rhine, viz. Gammarus tigrinus, Echinogammarus ischnus and Dikerogammarus villosus were tested. In single-species experiments, G. tigrinus and D. villosus showed similar preferences for a stony substratum, whereas E. ischnus mostly occupied the water column. The two-species aquarium experiments indicated direct interference competition for substratum and unequal mortality between G. tigrinus and D. villosus, with D. villosus being the stronger competitor. Competitive stress was influenced by population density, was size-dependent and varied between the different types of substratum due to substratum choice. G. tigrinus did not show any behaviour indicative of interference competition in the presence of E. ischnus, and neither did E. ischnus or D. villosus in the presence of any of the other gammarideans. Swimming in the water layer may already enable E. ischnus to minimise its encounters with the stone-dwelling D. villosus and G. tigrinus. To maximise the encounters between E. ischnus and D. villosus, a fish (Lepomis gibbosus) was added to occupy the water layer during the aquarium experiments. E. ischnus showed a higher mortality in the presence of both D. villosus and fish, probably due to increased stress, as shelter opportunities to escape the predators had been minimised. The study shows that interference competition between gammaridean species can explain the replacement of the North American invader G. tigrinus by D. villosus in the river Rhine. E. ischnus and D. villosus both Ponto-Caspian invaders did not show interference competition in our experiments and co-exist in the Rhine.

To conquer and persist : colonization and population development of the Ponto-Caspian amphipods Dikerogammarus villosus and Chelicorophium curvispinum on bare stone substrate in the main channel of the River Rhine

Macroinvertebrate communities on the stones in the Rhine are dominated by the Ponto-Caspian amphipods Chelicorophium curvispinum (since 1987) and Dikerogammarus villosus (since 1995), which have invaded the Rhine through canals connecting the large rivers of Europe. Colonization of bare stones suspended in the water of the Rhine main channel was studied. At the same time the macroinvertebrates drifting in the water layer were sampled. Macroinvertebrate populations on the newly colonized stones were followed for two months (June - August 2002). Bare stones were colonized from the water layer, with D. villosus and C. curvispinum most numerous from the start. Species richness was highest after one month. D. villosus and C. curvispinum continued to dominate the macroinvertebrate community on the stones throughout the experiment, representing 70-95 % of the total number of macroinvertebrates. In the first period week of colonization, especially juveniles of both amphipod species settled on the bare stones. After one week, the number of adults of D. villosus increased. After one month, ovigerous females of D. villosus became abundant on the newly colonized substrate. The numbers of adult C. curvispinum increased after one month and ovigerous females were present after two months. Newly settled populations resembled the amphipod populations present in the water layer, but started to deviate as colonization time increased, indicating that development of populations on stones became increasingly autonomous and less dependent on new colonization by amphipods from the water layer. Ovigerous females of both amphipods were much more abundant on the stones than in the water layer. Juvenile C. curvispinum were smaller on the stones than in the water layer, indicating that the stone substrate is important for reproduction of at least C. curvispinum. Most juveniles of this species first grow to a certain body length on the stones before they start drifting off and swimming in the water layer. C. curvispinum and D. villosus densities were positively correlated in the early stages of colonization, but showed an inverse relationship after longer colonisation time. As both dominant amphipods colonize new stone substrate from the water layer where they drift or swim, they may not depend on extra vectors, such as shipping, for dispersal through the connecting canals and within the river.