Aggressive encounters occur between competitors (particularly males) throughout the animal kingdom, and in some species can result in severe injury and death. Here we describe for the first time lethal interactions between male nematodes and provide evidence that the expression of this behaviour is developmentally controlled. Males of the entomopathogenic nematode Steinernema longicaudum coil around each other, resulting in injuries, paralysis and frequently death. The probability of death occurring between pairs of males was affected by the developmental pathway followed, being much greater among males that had passed through the infective juvenile (IJ, or dauer) stage than among males that had not. Post-IJ males are found only in newly colonised hosts, typically with few competing males present. Killing those few competitors may secure valuable resources (both females and a host cadaver for nourishment of offspring). Non-IJ males develop in subsequent generations within a host cadaver, where the presence of many closely related male competitors increases the risk:benefit ratio of fighting. Thus, passage through the IJ stage primes males for enhanced aggression in circumstances where this is more likely to result in increased reproductive success. Fighting occurred between males developing in mixed-sex social groups, indicating that it is an evolved trait and not an abnormal response to absence of females. This is supported by finding high mortality of males, but not of females, across a range of population densities in insect cadavers. We propose that these nematodes, with their relatively simple organization, may be a useful model for studies of aggression.

With the construction of wind farms all across the North Sea, numerous artificial reefs are created. These windmill artificial reefs (WARs) harbour high abundances of fish species which can be attracted from elsewhere or can be the result of extra production induced by these wind farms. To resolve the attraction-production debate in suddenly altered ecosystems (cf. wind farms), the possible consequences of attraction should be assessed; thereby bearing in mind that ecological traps may arise. In this paper we investigated whether the wind farms in the Belgian part of the North Sea act as ecological traps for pouting and Atlantic cod. Length-at-age, condition and diet composition of fish present at the windmill artificial reefs was compared to local and regional sandy areas. Fish data from the period 2009-2012 were evaluated. Mainly I- and II-group Atlantic cod were present around the WARs; while the 0- and I-group dominated for pouting. For Atlantic cod, no differences in length were observed between sites, indicating that fitness was comparable at the WARs and in sandy areas. No significant differences in condition index were observed for pouting. At the WARs, they were slightly larger and stomach fullness was enhanced compared to the surrounding sandy areas. Also diet differed considerably among the sites. The outcome of the proxies indicate that fitness of pouting was slightly enhanced compared to the surrounding sandy areas. No evidence was obtained supporting the hypothesis that the WARs act as an ecological trap for Atlantic cod and pouting.

a b s t r a c tIn experimentally infected insects, the sex ratio of first generation nematodes of five species of Steinernema was female-biased (male proportion 0.35-0.47). There was a similar female bias when the worms developed in vitro (0.37-0.44), indicating that the bias in these species is not due to a lower rate of infection by male infective juveniles (IJs). Experimental conditions influenced the proportion of males establishing in insects, indicating that male and female IJs differ in their behaviour. However, there was no evidence that males are the colonising sex in any species, contrary to what has previously been proposed. Time of emergence from the host in which the nematodes had developed influenced sex ratios in experimental infections. In three species (Steinernema longicaudum, Steinernema glaseri and Steinernema kraussei), early emerged nematodes had a higher proportion of males than those that emerged later, with the reverse trend for Steinernema carpocapsae and Steinernema feltiae. In a more detailed in vitro study of S. longicaudum, the proportion of males was similar whether or not the nematodes passed through the developmentally arrested IJ stage, indicating that the female bias is not due to failure of males to exit this stage. The sex ratio in vitro was independent of survival rate from juvenile to adult, and was female-biased even when all juveniles developed, indicating that the bias is not explained by failure of males to develop to adults. The female-biased sex ratio characteristic of Steinernema populations appears to be present from at least the early juvenile stage. We hypothesise that the observed female bias is the population optimal sex ratio, a response to cycles of local mate competition experienced by nematodes reproducing within insect hosts interspersed with periods of outbreeding with less closely related worms following dispersal. Ó

a b s t r a c tThere is evidence of competition within and between helminth species, but the mechanisms involved are not well described. In interference competition, organisms prevent each other from using the contested resource through direct negative interactions, either chemical or physical. Steinernema spp. are entomopathogenic nematodes; they enter a living insect host which they kill and consume with the aid of symbiotic bacteria. Several studies have demonstrated intra-and interspecific competition in Steinernema, mediated by a scramble for resources and by incompatibility of the bacterial symbiont. Here we describe a mechanism by which male Steinernema may compete directly for resources, both food (host) and females, by physically injuring or killing members of another species as well as males of their own species. A series of experiments was conducted in hanging drops of insect haemolymph. Males of each of four species (Steinernema longicaudum, Steinernema carpocapsae, Steinernema kraussei and Steinernema feltiae), representing three of the five phylogenetic clades of the genus, killed each other. Within 48 h, up to 86% of pairs included at least one dead male, compared with negligible mortality in single male controls. There was evidence of intraspecific difference: one strain of S. feltiae (4CFMO) killed while another (UK76) did not. Males also killed both females and males of other Steinernema spp. There was evidence of a hierarchy of killing, with highest mortality due to S. longicaudum followed by S. carpocapsae, S. kraussei and S. feltiae. Wax moth larvae were co-infected with members of two Steinernema spp. to confirm that killing also takes place in the natural environment of an insect cadaver. When insects were co-infected with one infective juvenile of each species, S. longicaudum males killed both S. feltiae UK76 and Steinernema hermaphroditum. Wax moths co-infected with larger, equal numbers of S. longicaudum and S. feltiae UK76 produced mainly S. longicaudum progeny, as expected based on hanging drop experiments. Ó

Objective and subjective components of resource value in lethal fights between male entomopathogenic nematodes ABSTRACT Males sometimes engage in fights over contested resources such as access to mates; in this case, fighting behaviour may be adjusted based on the value they place on the females.Resource value RV can have two components. Males can assess the quality of females, which constitutes an objective assessment of RV. Internal state such as previous mating experience can also influence motivation to fight thus constituting a subjective assessment of RV. If mating opportunities are scarce and available females have a major impact on the lifetime reproductive success of males, then fighting can be fatal; in this situation it is uncertain whether males would adjust fighting behaviour based on RV. We found that both female quality i.e., virginity (objective component of RV) and male mating status (subjective component of RV) influence fighting intensity between males of the entomopathogenic nematode Steinernema longicaudum which engage in lethal fights. Male nematodes were more likely to engage in fighting and fought longer and more frequently in the presence of virgin (high quality) females than in the presence of mated (lower quality) females. Male mating status was also found to influence fighting behaviour; mated males were the winners in staged fights between mated and virgin males. Mated males may have superior fighting ability (enhanced resource holding potential RHP), but RV asymmetries between mated and virgin males cannot be excluded. Males were more likely to win when they were resident, but we did not find a significant interaction effect between male mating and residency status.