Jean-Nicolas Beisel

Effect of gender on physiological and behavioural responses of Gammarus roeseli (Crustacea Amphipoda) to salinity and temperature

The importance of potentially interacting factors in organisms responses to a stress are often ignored or underestimated in ecotoxicology. In laboratory experiments we investigated how gender, temperature and age influence the behaviour and the physiology of the freshwater amphipod Gammarus roeseli under salinity stress. Our results revealed a significant higher sensitivity of females in survival, ventilation and ionoregulation whereas no inter-age differences were reported. Water temperature also exerted a significant effect in survival and ventilation of G. roeseli. Some of those factors appeared to interact significantly. This study provides evidence that gender can affect organisms responses to a stressor and consequently has to be considered while assessing a stress impact. We discussed the potential relationships between biological and behavioural responses.

Testing the potential of a ribosomal 16S marker for DNA metabarcoding of insects

Cytochrome c oxidase I (COI) is a powerful marker for DNA barcoding of animals, with good taxonomic resolution and a large reference database. However, when used for DNA metabarcoding, estimation of taxa abundances and species detection are limited due to primer bias caused by highly variable primer binding sites across the COI gene. Therefore, we explored the ability of the 16S ribosomal DNA gene as an alternative metabarcoding marker for species level assessments. Ten bulk samples, each containing equal amounts of tissue from 52 freshwater invertebrate taxa, were sequenced with the Illumina NextSeq 500 system. The 16S primers amplified three more insect species than the Folmer COI primers and amplified more equally, probably due to decreased primer bias. Estimation of biomass might be less biased with 16S than with COI, although variation in read abundances of two orders of magnitudes is still observed. According to these results, the marker choice depends on the scientific question. If the goal is to obtain a taxonomic identification at the species level, then COI is more appropriate due to established reference databases and known taxonomic resolution of this marker, knowing that a greater proportion of insects will be missed using COI Folmer primers. If the goal is to obtain a more comprehensive survey the 16S marker, which requires building a local reference database, or optimised degenerated COI primers could be more appropriate.

A manipulative parasite increasing an antipredator response decreases its vulnerability to a nonhost predator

Trophically transmitted parasites have to deal with the antipredator adaptations of their intermediate hosts. Some of these parasites induce behavioural changes in their intermediate hosts that make them more vulnerable to predation by definitive hosts. However, the adaptiveness of behavioural manipulation also depends on the probability of being eaten by a nonhost predator. Parasites might therefore try to use specific antipredator responses of intermediate hosts to avoid this dead end. We tested this hypothesis using the acanthocephalan Polymorphus minutus and its intermediate amphipod host, Gammarus roeseli. In their natural habitat, uninfected G. roeseli shelter near the river bottom while infected gammarids tend to cling onto floating materials. In microcosm experiments, infected amphipods were preyed upon by sticklebacks (nonhost predators) less than uninfected individuals when refuges were provided near the water surface. Regardless of their infection status, G. roeseli reacted to olfactory cues of predation by spending more time on refuges and near the surface. However, these behaviours were increased in infected gammarids. These results suggest that, in addition to specific induced behaviours, parasites may also enhance an existing antipredator behaviour to increase their own survival.

An acanthocephalan parasite boosts the escape performance of its intermediate host facing non-host predators

Among the potential effects of parasitism on host condition, the increased host abilities hypothesis is a counterintuitive pattern which might be predicted in complex-life-cycle parasites. In the case of trophic transmission, a parasite increasing its intermediate hosts performance facing non-host predators improves its probability of transmission to an adequate, definitive host. In the present study, we investigated the cost of infection with the acanthocephalan Polymorphus minutus on the locomotor/escape performance of its intermediate host, the crustacean Gammarus roeseli. This parasite alters the behaviour of its intermediate host making it more vulnerable to predation by avian definitive hosts. We assessed the swimming speeds of gammarids using a stressful treatment and their escape abilities under predation pressure. Despite the encystment of P. minutus in the abdomen of its intermediate host, infected amphipods had significantly higher swimming speeds than uninfected ones (increases of up to 35%). Furthermore, when interacting with the non-host crustacean predator Dikerogammarus villosus, the highest escape speeds and greatest distances covered by invertebrates were observed for parasitized animals. The altered behaviour observed among the manipulated invertebrates supported the increased host abilities hypothesis, which has until now remained untested experimentally. The tactic of increasing the ability of infected intermediate hosts to evade potential predation attempts by non-host species is discussed.

Effects of Sublethal Cadmium Exposure on Antipredator Behavioural and Antitoxic Responses in the Invasive Amphipod Dikerogammarus villosus

Amphipods are recognised as an important component of freshwater ecosystems and are frequently used as an ecotoxicological test species. Despite this double interest, there is still a lack of information concerning toxic impacts on ecologically relevant behaviours. The present study investigated the influence of cadmium (Cd), a non-essential heavy metal, on both antipredator behaviours and antitoxic responses in the invasive amphipod Dikerogammarus villosus under laboratory conditions. Amphipod behaviour (i.e. refuge use, aggregation with conspecifics, exploration and mobility) was recorded following a 4-min test-exposure to 500 µg Cd/L with or without a 24-h Cd pre-exposure and in the presence or absence of a high perceived risk of predation (i.e. water scented by fish predators and injured conspecifics). Following behavioural tests, malondialdehyde (MDA) levels, a biomarker for toxic effect, and energy reserves (i.e. lipid and glycogen contents) were assessed. Cd exposures induced (1) cell damage reflected by high MDA levels, (2) erratic behaviour quantified by decreasing refuge use and exploration, and increasing mobility, and (3) a depletion in energy reserves. No significant differences were observed between 4-min test-exposed and 24-h pre-exposed individuals. Gammarids exposed to Cd had a disturbed perception of the alarm stimuli, reflected by increased time spent outside of refuges and higher mobility compared to gammarids exposed to unpolluted water. Our results suggest that Cd exposure rapidly disrupts the normal behavioural responses of gammarids to alarm substances and alters predator-avoidance strategies, which could have potential impacts on aquatic communities.

Patterns of Fluctuating Asymmetry and Shape Variation in Chironomus riparius (Diptera, Chironomidae) Exposed to Nonylphenol or Lead

Deformities and fluctuating asymmetry in chironomid larvae have been proposed as sensitive indicators of biological stress and are commonly used to assess the ecological impact of human activities. In particular, they have been associated in Chironomus riparius, the most commonly used species, with heavy metal and pesticide river pollution. In this study, the effect of lead and 4-nonylphenol on mouthpart morphological variation of Chironomus riparius larvae was investigated by traditional and geometric morphometrics. For this purpose, first to fourth instar larvae were exposed to sediment spiked with lead (from 3.0 to 456.9 mg/kg dry weight) or 4-NP (from 0.1 to 198.8 mg/kg dry weight). Mentum phenotypic response to pollutants was assessed by four parameters: (1) the frequency of deformities, (2) fluctuating asymmetry of mentum length, (3) fluctuating asymmetry of mentum shape and (4) the mentum mean shape changes. Despite the bioaccumulation of pollutants in the chironomid’s body, no significant differences between control and stressed groups were found for mouthpart deformities and fluctuating asymmetry of mentum length. Slight effects on mentum shape fluctuating asymmetry were observed for two stressed groups. Significant mean shape changes, consisting of tooth size increase and tooth closing, were detected for lead and 4-NP exposure respectively. Those variations, however, were negligible in comparison to mentum shape changes due to genetic effects. These results suggest that the use of mentum variation as an indicator of toxic stress in Chironomus riparius should be considered cautiously.

Behavioural and Physiological Responses of Gammarus pulex Exposed to Cadmium and Arsenate at Three Temperatures: Individual and Combined Effects

This study aimed at investigating both the individual and combined effects of cadmium (Cd) and arsenate (AsV) on the physiology and behaviour of the Crustacean Gammarus pulex at three temperatures (5, 10 and15°C). G. pulex was exposed during 96 h to (i) two [Cd] alone, (ii) two [AsV] alone, and (iii) four combinations of [Cd] and [AsV] to obtain a complete factorial plane. After exposure, survival, [AsV] or [Cd] in body tissues, behavioural (ventilatory and locomotor activities) and physiological responses (iono-regulation of [Na+] and [Cl−] in haemolymph) were examined. The interactive effects (antagonistic, additive or synergistic) of binary mixtures were evaluated for each tested temperature using a predictive model for the theoretically expected interactive effect of chemicals. In single metal exposure, both the internal metal concentration in body tissues and the mortality rate increased along metallic gradient concentration. Cd alone significantly impaired both [Na+] and [Cl−] while AsV alone had a weak impact only on [Cl−]. The behavioural responses of G. pulex declined with increasing metal concentration suggesting a reallocation of energy from behavioural responses to maintenance functions. The interaction between AsV and Cd was considered as ‘additive’ for all the tested binary mixtures and temperatures (except for the lowest combination at 10°C considered as “antagonistic”). In binary mixtures, the decrease in both ventilatory and locomotor activities and the decline in haemolymphatic [Cl−] were amplified when respectively compared to those observed with the same concentrations of AsV or Cd alone. However, the presence of AsV decreased the haemolymphatic [Na+] loss when G. pulex was exposed to the lowest Cd concentration. Finally, the observed physiological and behavioural effects (except ventilation) in G. pulex exposed to AsV and/or Cd were exacerbated under the highest temperature. The discussion encompasses both the toxicity mechanisms of these metals and their interaction with rising temperature.

Parasite-induced changes in the diet of a freshwater amphipod: field and laboratory evidence

Trophically transmitted parasites are likely to strongly influence food web-structure. The extent to which they change the trophic ecology of their host remains nevertheless poorly investigated and field evidence is lacking. This is particularly true for acanthocephalan parasites whose invertebrate hosts can prey on other invertebrates and contribute to leaf-litter breakdown. We used a multiple approach combining feeding experiments, neutral lipids and stable isotopes to investigate the trophic ecology of the freshwater amphipod Gammarus roeseli parasitized by the bird acanthocephalan Polymorphus minutus. Infected compared to uninfected amphipods consumed as many dead isopods, but fewer live isopods and less leaf material. Infection had no influence on the total concentration of neutral lipids. Contrary to what we expected based on laboratory findings, the nitrogen isotope signature, which allows for the estimation of consumers trophic position, was not influenced by infection status. Conversely, the carbon isotope signature, which is used to identify food sources, changed with infection and suggested that the diet of infected G. roeseli includes less perilithon (i.e. fixed algae on rocks, stones) but more terrestrial inputs (e.g. leaf material) than that of uninfected conspecifics. This study shows evidence of changes in the trophic ecology of P. minutus-infected G. roeseli and we stress the need to complement feeding experiments with field data when investigating top-down effects of infection in an opportunistic feeder which adapts its diet to the available food sources.

Geographic patterns in freshwater gammarid invasions: an analysis at the pan-European scale

Abstract.Biogeographic characteristics of freshwater gammarids at a pan-European scale were investigated to describe spatial gradients of richness and their relationships with the pattern of invasive species distribution. Species richness appeared to be linked to latitude and among the 25 European biogeographical areas, the Ponto-Caspian basin exhibited a particular position, being by far the most diverse area. A link between the species richness of an area and the number of invasive species coming from this area was observed, showing that rich areas are more prone to shelter potential invaders than others. The patterns of gammarid dispersals among European waterways suggest that, rather than a random distribution of invaders, a clump occurred in basins which served as refuge areas during the last glaciations. Several reasons explaining the pattern are discussed, including the possibility that among species evolving in a stimulating environment some of them have had the opportunity to develop special competitive attributes.

Acanthocephalan parasites: help or burden in gammarid amphipods exposed to cadmium?

We investigated the influence of the acanthocephalan parasite Polymorphus minutus on the mortality of its intermediate host, Gammarus roeseli, exposed to cadmium, by the measure of LC50–96h values as well as the bioaccumulation of cadmium both in the host and in its parasite. LC50 results revealed that infected G. roeseli males died less under cadmium stress than uninfected ones; while the converse has been observed in females. Cadmium resistance of infected males could be explained by a weaker bioconcentration factor (BCF) than in females. The lower BCF in infected individuals was closely related with an uptake of cadmium by P. minutus in its host. Nevertheless, although infected females had both weaker BCF and cadmium concentration in their body, the presence of P. minutus did not induce lower mortality than uninfected females. On the contrary, their sensitivity to cadmium was increased by the presence of P. minutus. We discuss the hypothesis that differences of mortality between uninfected and infected gammarids could be explained by a difference of cadmium bioconcentration in host, and by the cadmium bioaccumulation in the parasite. Indeed, results suggested that P. minutus could help G. roeseli to face with stress, what contributed to keep the host alive and favour the parasite transmission.