Establishing arthropod community composition using metabarcoding: Surprising inconsistencies between soil samples and preservative ethanol and homogenate from Malaise trap catches.

DNA metabarcoding allows the analysis of insect communities faster and more efficiently than ever before. However, metabarcoding can be conducted through several approaches, and the consistency of results across methods has rarely been studied. We compare the results obtained by DNA metabarcoding of the same communities using two different markers - COI and 16S - and three different sampling methods: (a) homogenized Malaise trap samples (homogenate), (b) preservative ethanol from the same samples, and (c) soil samples. Our results indicate that COI and 16S offer partly complementary information on Malaise trap samples, with each marker detecting a significant number of species not detected by the other. Different sampling methods offer highly divergent estimates of community composition. The community recovered from preservative ethanol of Malaise trap samples is significantly different from that recovered from homogenate. Small and weakly sclerotized insects tend to be overrepresented in ethanol while strong and large taxa are overrepresented in homogenate. For soil samples, highly degenerate COI primers pick up large amounts of nontarget DNA and only 16S provides adequate analyses of insect diversity. However, even with 16S, very little overlap in molecular operational taxonomic unit (MOTU) content was found between the trap and the soil samples. Our results demonstrate that none of the tested sampling approaches is satisfactory on its own. For instance, DNA extraction from preservative ethanol is not a valid replacement for destructive bulk extraction but a complement. In future metabarcoding studies, both should ideally be used together to achieve comprehensive representation of the target community.

Complementary methods assessing short and long-term prey of a marine top predator ‒ Application to the grey seal-fishery conflict in the Baltic Sea.

The growing grey seal (Halichoerus grypus) population in the Baltic Sea has created conflicts with local fisheries, comparable to similar emerging problems worldwide. Adequate information on the foraging habits is a requirement for responsible management of the seal population. We investigated the applicability of available dietary assessment methods by comparing morphological analysis and DNA metabarcoding of gut contents (short-term diet; n = 129/125 seals, respectively), and tissue chemical markers i.e. fatty acid (FA) profiles of blubber and stable isotopes (SIs) of liver and muscle (mid- or long-term diet; n = 108 seals for the FA and SI markers). The methods provided complementary information. Short-term methods indicated prey species and revealed dietary differences between age groups and areas but for limited time period. In the central Baltic, herring was the main prey, while in the Gulf of Finland percid and cyprinid species together comprised the largest part of the diet. Perch was also an important prey in the western Baltic Proper. The DNA analysis provided firm identification of many prey species, which were neglected or identified only at species group level by morphological analysis. Liver SIs distinguished spatial foraging patterns and identified potentially migrated individuals, whereas blubber FAs distinguished individuals frequently utilizing certain types of prey. Tissue chemical markers of adult males suggested specialized feeding to certain areas and prey, which suggest that these individuals are especially prone to cause economic losses for fisheries. We recommend combined analyses of gut contents and tissue chemical markers as dietary monitoring methodology of aquatic top predators to support an optimal ecosystem-based management.

Seasonally varying marine influences on the coastal ecosystem detected through molecular gut analysis.

Terrestrial predators on marine shores benefit from the inflow of organisms and matter from the marine ecosystem, often causing very high predator densities and indirectly affecting the abundance of other prey species on shores. This indirect effect may be particularly strong if predators shift diets between seasons. We therefore quantified the seasonal variation in diet of two wolf spider species that dominate the shoreline predator community, using molecular gut content analyses with general primers to detect the full prey range. Across the season, spider diets changed, with predominantly terrestrial prey from May until July and predominantly marine prey (mainly chironomids) from August until October. This pattern coincided with a change in the spider age and size structure, and prey abundance data and resource selection analyses suggest that the higher consumption of chironomids during autumn is due to an ontogenetic diet shift rather than to variation in prey abundance. The analyses suggested that small dipterans with a weak flight capacity, such as Chironomidae, Sphaeroceridae, Scatopsidae and Ephydridae, were overrepresented in the gut of small juvenile spiders during autumn, whereas larger, more robust prey, such as Lepidoptera, Anthomyidae and Dolichopodidae, were overrepresented in the diet of adult spiders during spring. The effect of the inflow may be that the survival and growth of juvenile spiders is higher in areas with high chironomid abundances, leading to higher densities of adult spiders and higher predation rates on the terrestrial prey next spring.

Effect of repeated exposure to Plasmodium relictum (lineage SGS1) on infection dynamics in domestic canaries.

Parasites are known to exert strong selection pressures on their hosts and, as such, favour the evolution of defence mechanisms. The negative impact of parasites on their host can have substantial consequences in terms of population persistence and the epidemiology of the infection. In natural populations, however, it is difficult to assess the cost of infection while controlling for other potentially confounding factors. For instance, individuals are repeatedly exposed to a variety of parasite strains, some of which can elicit immunological memory, further protecting the host from subsequent infections. Cost of infection is, therefore, expected to be particularly strong for primary infections and to decrease for individuals surviving the first infectious episode that are re-exposed to the pathogen. We tested this hypothesis experimentally using avian malaria parasites (Plasmodium relictum-lineage SGS1) and domestic canaries (Serinus canaria) as a model. Hosts were infected with a controlled dose of P. relictum as a primary infection and control birds were injected with non-infected blood. The changes in haematocrit and body mass were monitored during a 20 day period. A protein of the acute phase response (haptoglobin) was assessed as a marker of the inflammatory response mounted in response to the infection. Parasite intensity was also monitored. Surviving birds were then re-infected 37 days post primary infection. In agreement with the predictions, we found that primary infected birds paid a substantially higher cost in terms of infection-induced reduction in haematocrit compared with re-exposed birds. After the secondary infection, re-exposed hosts were also able to clear the infection at a faster rate than after the primary infection. These results have potential consequences for the epidemiology of avian malaria, since birds re-exposed to the pathogen can maintain parasitemia with low fitness costs, allowing the persistence of the pathogen within the host population.

Distribución temporal de aves rapaces diurnas en la reserva "Playón de Mismaloya", Jalisco, México / Temporal distribution of diurnal rapacious birds in \"Playón de Mismaloya\", Jalisco, México

Diurnal raptors were counted monthly in the "Playón de Mismaloya", reserve, Jalisco, Mexico, from November 1997 to October 1998. We identified 11 species; eight of them migrants, one resident and two resident with migrant populations. The highest increase in total number of individuals was from December to March, influenced mainly by the presence of Caracara polancus (Crested Caracara), Falco sparverius (American Kestrel) and Pandion haliaetus (Osprey). The two former species were observed frequently in open areas, where they could detect prey more easily, while ospreys were found only in estuaries, lagoons and beach, where food was available.