A Pretty Kettle of Fish: A Review on the Current Challenges in Mediterranean Teleost Reproduction.

The Mediterranean region is facing several environmental changes and pollution issues. Teleosts are particularly sensitive to these challenges due to their intricate reproductive biology and reliance on specific environmental cues for successful reproduction. Wild populations struggle with the triad of climate change, environmental contamination, and overfishing, which can deeply affect reproductive success and population dynamics. In farmed species, abiotic factors affecting reproduction are easier to control, whereas finding alternatives to conventional diets for farmed teleosts is crucial for enhancing broodstock health, reproductive success, and the sustainability of the aquaculture sector. Addressing these challenges involves ongoing research into formulating specialized diets, optimizing feeding strategies, and developing alternative and sustainable feed ingredients. To achieve a deeper comprehension of these challenges, studies employing model species have emerged as pivotal tools. These models offer advantages in understanding reproductive mechanisms due to their well-defined physiology, genetic tractability, and ease of manipulation. Yet, while providing invaluable insights, their applicability to diverse species remains constrained by inherent variations across taxa and oversimplification of complex environmental interactions, thus limiting the extrapolation of the scientific findings. Bridging these gaps necessitates multidisciplinary approaches, emphasizing conservation efforts for wild species and tailored nutritional strategies for aquaculture, thereby fostering sustainable teleost reproduction in the Mediterranean.

Increasing marine trophic web knowledge through DNA analyses of fish stomach content: a step towards an ecosystem-based approach to fisheries research.

Multispecies and ecosystem models, which are key for the implementation of ecosystem-based approaches to fisheries management, require extensive data on the trophic interactions between marine organisms, including changes over time. DNA metabarcoding, by allowing the simultaneous taxonomic identification of the community present in hundreds of samples, could be used for speeding up large-scale stomach content data collection. Yet, for DNA metabarcoding to be routinely implemented, technical challenges should be addressed, such as the potentially complicated sampling logistics, the detection of a high proportion of predator DNA, and the inability to provide reliable abundance estimations. Here, we present a DNA metabarcoding assay developed to examine the diet of five commercially important fish, which can be feasibly incorporated into routinary samplings. The method is devised to speed up the analysis process by avoiding the stomach dissection and content extraction steps, while preventing the amplification of predator DNA by using blocking primers. Tested in mock samples and in real stomach samples, the method has proven effective and shows great effectiveness discerning diet variations due to predator ecology or prey availability. Additionally, by applying our protocol to mackerel stomachs previously analyzed by visual inspection, we showcase how DNA metabarcoding could complement visually based data by detecting overlooked prey by the visual approach. We finally discuss how DNA metabarcoding-based data can contribute to trophic data collection. Our work reinforces the potential of DNA metabarcoding for the study and monitoring of fish trophic interactions and provides a basis for its incorporation into routine monitoring programs, which will be critical for the implementation of ecosystem-based approaches to fisheries management.

Effects of Dietary Vegetable Oil Mixtures including Soybean Oil on Intestinal Oxidative Stress in Gilthead Sea Bream (Sparus aurata).

Fish oil is commonly replaced by vegetable oils in sea bream diets, but little is known about their effects on intestinal health regarding oxidative stress biomarkers. The negative effects of lipid peroxidation on digestive mucosa could have consequences in animal nutrition and welfare. In this study, five isonitrogenous (46%) and isolipidic (22%) diets with 75% of vegetable oils inclusion were evaluated: soybean oil (S) alone or different mixtures containing soybean oil with linseed (SL), linseed and rapeseed (SLR), linseed and palm (SLP), and linseed, rapeseed, and palm (SLRP). Gilthead sea bream juveniles were fed twice a day for 18 weeks. Pyloric caeca and proximal intestine samples were collected 24 h post feeding for lipid peroxidation (LPO), antioxidant enzyme activities (SOD, CAT, GPx, GST, and GR) and gene expression analyses. Pyloric caeca presented larger unhealthy changes in oxidative status than proximal intestine. Although SL-fed fish showed the highest antioxidant activities, they were unable to cope with LPO that in pyloric caeca was 31.4 times higher than in the other groups. Instead, SLP fish presented the best oxidative status, with low LPO levels, antioxidant enzyme activities, and gene expression. In summary, between the vegetable oils dietary mixtures tested, SPL would maintain better intestinal health.

Food web ecology of Gulf Stream flounder (Citharichthys arctifrons): a continental shelf perspective.

Gulf Stream flounder, Citharichthys arctifrons, are regularly observed in fish diets of the northeast U.S. continental shelf, yet lack commercial value and are often ignored. Similarly, Gulf Stream flounder diets of the Northwest Atlantic have remained largely unexamined, except for a brief period from 1976 to 1980. To better understand their role in the ecosystem, juvenile through adult Gulf Stream flounder were examined both as a predator and prey, and the magnitude of their feeding footprint (removal of prey biomass) was quantified for the northeast U.S. continental shelf. Their stomachs were sampled from 2005 to 2010, with the majority examined in the field macroscopically. Due to large proportions of unidentifiable prey, the effort was expanded in 2011-2012, and all stomachs were processed in the laboratory microscopically. Gulf Stream flounder were consumed by 15 fish, and what they eat (percentage mass and percentage frequency of occurrence) was documented by season, spatial region and year. Highly benthivorous, Gammaridea and Polychaeta dominated the diet in all years, seasons and regions, but Ophiuroidea (brittle stars) were also prominent in Southern New England during the spring. Gulf Stream flounder diets remained consistent across regions and time, with only a few feeding differences between seasons and one region. Relative to the productivity of benthos for this shelf ecosystem, the feeding footprint of Gulf Stream flounder was minor for their predominant benthic prey with a maximum percentage of benthos production eaten of 0.01% m-2 in Southern New England. With an ecosystem perspective, this feeding information offers a foundation for improving fisheries management among shared living marine resources considering benthic habitat and prey availability.

Background per- and polyfluoroalkyl substances (PFAS) in laboratory fish diet: Implications for zebrafish toxicological studies.

Current attention is focused on determining the potential for per- and polyfluoroalkyl substances (PFAS) to adversely impact human health. Zebrafish are a popular biological model because they share early development pathways with humans. A dietary exposure paradigm is growing in popularity in the zebrafish model because the outcomes often translate to humans. To create a diet of known composition, it is crucial to understand background PFAS levels present in zebrafish diet. Background PFAS, if present, potentially confounds interpretation of toxicological data. To date, no studies document the PFAS background levels in laboratory fish diet and there is only limited information on some pet foods. The objective of this study was to develop and validate an analytical method for up to 50 target PFAS in high lipid and protein content laboratory fish diets and pet foods. Long-chain perfluoroalkyl carboxylic acids (C9-C13) and perfluorooctane sulfonate (PFOS) were quantified in 11 out of 16 laboratory fish diets and in three out of five pet fish foods. Foods for pet birds, lizards, and dogs were below the limit of detection for all PFAS. In two of the laboratory fish diets, PFOS concentrations were >1.3 ng/g and the total PFAS for the three laboratory fish diets exceeded 1.0 ng/g. Hundreds of biomedical laboratories across the world utilize these commercial laboratory fish diets, and these results indicate that numerous zebrafish colonies may be inadvertently receiving significant dietary PFAS exposures. In light of this new information, it is critical to design PFAS studies with appropriate controls with measured background PFAS concentrations in the diet and to urge caution when interpreting the results.

Alternative Proteins for Fish Diets: Implications beyond Growth.

Aquaculture has been challenged to find alternative ingredients to develop innovative feed formulations that foster a sustainable future growth. Given the most recent trends in fish feed formulation on the use of alternative protein sources to decrease the dependency of fishmeal, it is fundamental to evaluate the implications of this new paradigm for fish health and welfare. This work intends to comprehensively review the impacts of alternative and novel dietary protein sources on fish gut microbiota and health, stress and immune responses, disease resistance, and antioxidant capacity. The research results indicate that alternative protein sources, such as terrestrial plant proteins, rendered animal by-products, insect meals, micro- and macroalgae, and single cell proteins (e.g., yeasts), may negatively impact gut microbiota and health, thus affecting immune and stress responses. Nevertheless, some of the novel protein sources, such as insects and algae meals, have functional properties and may exert an immunostimulatory activity. Further research on the effects of novel protein sources, beyond growth, is clearly needed. The information gathered here is of utmost importance, in order to develop innovative diets that guarantee the production of healthy fish with high quality standards and optimised welfare conditions, thus contributing to a sustainable growth of the aquaculture industry.

The inverse association between a fish consumption biomarker and gingival inflammation and periodontitis: A population-based study.

AIM: The metabolite 3-carboxy-4-methyl-5-propyl-2-furanpropionic acid (CMPF) is a fatty fish-intake biomarker. We investigated the association between plasma levels of CMPF in relation to gingival inflammation and periodontitis case definition, as well as the extent and severity variables. MATERIALS AND METHODS: The Malmö Offspring Study is a population-based study, and the Malmö Offspring Dental Study (MODS) is its dental arm, including periodontal charting. Plasma CMPF was measured using liquid chromatography-mass spectrometry and studied in relation to periodontal diagnosis and parameters using multivariable linear or logistic regression modelling adjusting for age, sex, education, body mass index, fasting glucose, and smoking. RESULTS: Metabolite data were available for 922 MODS participants. Higher CMPF levels were associated with less gingival inflammation (ß = -2.12, p = .002) and lower odds of severe periodontitis (odds ratio [OR] = 0.74, 95% confidence interval [CI]: 0.56 to 0.98). Higher CMPF levels were also associated with more teeth (ß = 0.19, p = .001), lower number of periodontal pockets (≥4 mm) (ß = -1.07, p = .007), and lower odds of having two or more periodontal pockets of ≥6 mm (OR = 0.80, 95% CI: 0.65 to 0.98) in fully adjusted models. CONCLUSIONS: CMPF, a validated biomarker of fatty fish consumption, is associated with less periodontal inflammation and periodontitis. Residual confounding cannot be ruled out, and future studies are warranted.

Effect of replacing fish meal in fish diet with shrimp by-product meal on growth performance, feed utilization, length-weight relationship and condition factors of Nile tilapia, Oreochromis niloticus (Linnaeus, 1758) / Efecto de reemplazar la harina de pescado en la dieta de pescado por harina de subproducto de camarón sobre el rendimiento del crecimiento, la utilización del alimento, la relación longitud-peso y los factores de condición de la tilapia del Nilo, Oreochromis niloticus (Linnaeus, 1758)

SUMMARY: The present work aimed to study the effect of replacing fish meal (FM) in the fish diet with shrimp by-product meal (SBM) on the growth performance parameters of the Nile Tilapia, Oreochromis niloticus. A total of 200 specimens of monosex O. niloticus fries were obtained from a private fish farm at Kafr El-Sheikh Governorate. They were transported to the fish laboratory at the Animal House of Zoology Department, Faculty of Science, Al-Azhar University. After two weeks for acclimation, the fish were divided into 5 groups, 2 replicates for each group (20 fish in each replicate). The five experimental diets were: C: control group with20 % fish meal (FM) and 0 % shrimp by-product meal (SM); T1, T2, T3, and T4 FM was replaced with SBM as 25 %, 50 %, 75 % and 100 %, respectively. Results indicated that the highest final length, growth in length, length gain, daily length gain and other growth performance parameters including the relationship between length and weight and condition factors of O. niloticus were recorded in T4 group (fed diet in which fish meal was completely replaced with shrimp by-product meal); while, their lowest values were recorded in control group. On the other hand, feed utilization parameters (feed intake, food conversion ratio, the maximum values of feed efficiency ratio and protein efficiency ratio) were recorded in T4 group and the minimum values were recorded in T3 group (fed diet in which 75 % of fish meal was replaced with shrimp by-product meal).

RESUMEN: El presente trabajo tuvo como objetivo estudiar el efecto de sustituir la harina de pescado (HP) en la dieta del pescado por harina de subproductos de camarón (HSC) sobre los parámetros de rendimiento de crecimiento de la Tilapia del Nilo, Oreochromis niloticus. Se obtuvieron un total de 200 especímenes de monosexo de O. niloticus de una piscifactoría privada en la gobernación de Kafr El-Sheikh. Fueron transportados al laboratorio de peces en el Departamento de Zoología de la Facultad de Ciencias, de la Universidad Al-Azhar. Después de dos semanas de aclimatación, los peces se dividieron en 5 grupos: Se realizaron dos repeticiones para cada grupo (20 peces en cada repetición). Las cinco dietas experimentales fueron: C: grupo control con 20 % de harina de pescado (HP) y 0 % de harina de subproductos de camarón (HSC); T1, T2, T3 y T4 FM se reemplazó con HSC con 25 %, 50 %, 75 % y 100 %, respectivamente. Los resultados indicaron que la longitud final más alta, el crecimiento en longitud, la ganancia de longitud, la ganancia de longitud diaria y otros parámetros de rendimiento del crecimiento, como además la relación entre la longitud y el peso, y los factores de condición de O. niloticus, se registraron en el grupo T4 (con una dieta reemplazada con harina de subproducto de camarón); mientras que, sus valores más bajos se registraron en el grupo control. Por otro lado, los parámetros de utilización del alimento (ingesta de alimento, índice de conversión de alimento, los valores máximos de índice de eficiencia alimenticia e índice de eficiencia proteica) se registraron en el grupo T4 y los valores mínimos se registraron en el grupo T3 (alimentación con dieta en la que el 75 % de la harina de pescado fue reemplazada por harina de subproductos de camarón).

Phylogeny, body morphology, and trophic level shape intestinal traits in coral reef fishes.

Trait-based approaches are increasingly used to study species assemblages and understand ecosystem functioning. The strength of these approaches lies in the appropriate choice of functional traits that relate to the functions of interest. However, trait-function relationships are often supported by weak empirical evidence.Processes related to digestion and nutrient assimilation are particularly challenging to integrate into trait-based approaches. In fishes, intestinal length is commonly used to describe these functions. Although there is broad consensus concerning the relationship between fish intestinal length and diet, evolutionary and environmental forces have shaped a diversity of intestinal morphologies that is not captured by length alone.Focusing on coral reef fishes, we investigate how evolutionary history and ecology shape intestinal morphology. Using a large dataset encompassing 142 species across 31 families collected in French Polynesia, we test how phylogeny, body morphology, and diet relate to three intestinal morphological traits: intestinal length, diameter, and surface area.We demonstrate that phylogeny, body morphology, and trophic level explain most of the interspecific variability in fish intestinal morphology. Despite the high degree of phylogenetic conservatism, taxonomically unrelated herbivorous fishes exhibit similar intestinal morphology due to adaptive convergent evolution. Furthermore, we show that stomachless, durophagous species have the widest intestines to compensate for the lack of a stomach and allow passage of relatively large undigested food particles.Rather than traditionally applied metrics of intestinal length, intestinal surface area may be the most appropriate trait to characterize intestinal morphology in functional studies.

Assessment of Astaxanthin Accumulation in Hepatocytes of Atlantic Salmon Fed Different Diets Using NMR Spectroscopy.

This study aimed to assess the astaxanthin (Ax) accumulation in hepatocytes isolated from farmed Atlantic salmon fed different diets (rich marine, poor, poor with marine phospholipids (MPL) and poor with docosahexaenoic acid (DHA)). Nuclear magnetic resonance (NMR) spectroscopy was used for the Ax detection and quantification. The use of the 13C-enriched Ax allowed the assessment of short-time Ax metabolism. The substitution of fish oil and meal in fish feed on plant analogs and the addition of MPL caused further catabolism and decrease of Ax accumulation in hepatocytes from 17 to about 6 mg/kg or to almost zero in the case of DHA addition. Signals assignment of the native and 13C-enriched astaxanthin in acetone were performed using 1D and 2D NMR spectra.

Green macroalgae blooms (Ulva spp.) influence trophic ecology of juvenile flatfish differently in sandy beach nurseries.

Opportunistic green macroalgae blooms increasingly affect coastal areas worldwide. Understanding their impacts on organisms that use this zone, such as juvenile flatfish, is critical. By combining stable isotope data, digestive tract contents and community analyses of flatfish and their potential prey (benthic macroinvertebrates) from two North-East Atlantic sandy beaches (one impacted by blooms and one not), we detected similar and species-specific trophic changes among three co-occurring species (sand sole, plaice and turbot). Across flatfish species, juveniles displayed more opportunistic foraging behavior at the impacted site. Differently, plaice and sand sole relied more on the additional basal resource (Ulva spp.) than turbot. Finally, sand sole and turbot presented a stronger diet shift at the impacted site than plaice. We hypothesize that the species-specific response to the blooms are mostly driven by how the flatfish detect their prey (using visual and/or chemical cues) and when they forage (diurnal or nocturnal foraging).

Importance of ciliates as food for fish larvae in a shallow sea bay and a large shallow lake.

We estimated the consumption of planktonic ciliates by fish larvae in the Väinameri Sea (a shallow semi-enclosed bay of the Baltic Sea) and Lake Võrtsjärv (a shallow and eutrophic lake). Our primary hypothesis was that planktonic ciliates constitute a substantial component of the diet of fish larvae in both environments. We also assumed that the contribution of ciliates to larval nutrition is bigger in lacustrine than in marine environment because ciliates are usually more abundant in lakes. The nutrition of field collected larval fish was determined by gut content analysis using epifluorescence microscopy. Our study revealed that ciliates occurred in the alimentary tracts of all fish species examined. We discovered that the consumption of ciliates by first-feeding fish larvae contributed approximately 40 and 60% of their total consumed carbon in the Väinameri and in Võrtsjärv, respectively. Ciliates represent essentially important food for fish larvae and sufficient protozoan food may enhance larval growth in the beginning of the exogenous feeding and shorten the most vulnerable period in larval stage before shifting to larger prey.

Know your fish: A novel compound-specific isotope approach for tracing wild and farmed salmon.

The rapid expansion of the aquaculture industry with carnivorous fish such as salmon has been accompanied by an equally rapid development in alternative feed ingredients. This has outpaced the ability of prevailing authentication method to trace the diet and origins of salmon products at the retail end. To close this gap, we developed a new profiling tool based on amino acid δ13C fingerprints. With this tool, we discriminated with high-accuracy among wild-caught, organically, and conventionally farmed salmon groups, as well as salmon fed alternative diets such as insects and macroalgae. Substitution of fishmeal with macroalgae was detected at 5% difference level. The δ13C fingerprints of essential amino acids appear particularly well suited for tracing protein sources, and the non-essentials for tracing lipid origins (terrestrial vs. aquatic). In an industry constantly developing new feed proteins and functional additives, our method is a promising tool for tracing salmon and other seafood products.

10th European Zebrafish Meeting 2017, Budapest: Husbandry Workshop Summary.

A husbandry workshop on July 3, 2017, at the 10th European Zebrafish Meeting in Budapest, Hungary (July 3-July 7, 2017), focused on the standardization, optimization, and streamlining of fish facility procedures. Standardization can be achieved for example by developing novel software and hardware tools, such as a fish facility database for husbandry and environmental facility management (Zebrabase, Oltova), or a hand-held, air-pressurized fish feeder for consistent food distribution (Blowfish, Argenton). Streamlining is achieved when work hours are reduced, as with the standardized fish feeder, or by limiting the number and types of fish diets and observing the effect on animal welfare and performance (Barton). Testing the characteristics of new fish diets and observing whether they produce better experimental outcomes (Certal) optimizes diets and improves fish productivity. Collectively, the workshop presentations emphasized how consistency and harmonization of husbandry procedures within and across aquatic facilities yield reproducible scientific outcomes.

Application of DNA metabarcoding on faeces to identify European catfish Silurus glanis diet.

In this study, the results of conventional stomach-content analysis are compared with the recent DNA metabarcoding approach on faeces to identify fish species consumed by non-native European catfish Silurus glanis in the Garonne River (south-western France), with a special emphasis on anadromous prey. Fourteen prey species were identified in the stomach contents or faeces, including four anadromous fish species. Despite higher intestine than stomach emptiness, more species were identified through faecal analysis (11 of 14) than through stomach-content analysis (five of 14) suggesting that DNA metabarcoding on faeces is an efficient, non-intrusive technique to study the diet of predatory fishes.

Dietary administration of EDC mixtures: A focus on fish lipid metabolism.

Many man-made chemical compounds are recognized as endocrine disruptors and once released into the environment are likely to spread and bioaccumulate in wild species. Due to their lipophilic nature, these substances pass through the cell membrane or bind to specific receptors activating physiological responses that in the long run can cause reproductive impairment, physiological disorders, including the occurrence of metabolic syndromes. One significant source of contamination is represented by the consumption of polluted food. As a consequence, different environmental pollutants, with similar or different modes of action, can accumulate in organisms and biomagnify along the food web, finally targeting humans. The aim of this study was to analyze, under controlled conditions, the effects induced by the consumption of contaminated diets, focusing on the effects exerted at hepatic level. Juvenile seabream were fed for 21days a diet enriched with different combinations of pollutants, nonylphenol (NP), tert-octylphenol (t-OP) and bisphenol A (BPA). The different diets containing 5mg/kg bw of each contaminant, were formulated as follows: NP+tOP, BPA+NP, BPA+tOP and NP+BPA+tOP (NBO). EDCs, at the doses administered, showed low biomagnification factor (BMF), suggesting that these pollutants hardly accumulate in muscles. The results obtained at hepatic level pinpointed the steatotic effect of all the administered diets, associated to a modulation of the expression of genes involved in lipid metabolism (ppars, fas, lpl, and hsl). Results were compared to those obtained in previous studies in which fish were fed single pollutants evidencing that the administration of mixture of contaminants exerts a milder lipogenic effect, highlighting the contrasting/antagonistic interaction establishing among chemicals. Noteworthy was the setup of a new chromatographic method to detect the presence of the selected chemical in fish muscle and the application of Fourier Transform Infrared (FT-IR) analysis to evaluate pollutant-induced changes in the liver macromolecular building.

Trophic structure of a coastal fish community determined with diet and stable isotope analyses.

A combination of dietary guild analysis and nitrogen (δ(15) N) and carbon (δ(13) C) stable-isotope analysis was used to assess the trophic structure of the fish community in Rhode Island and Block Island Sounds, an area off southern New England identified for offshore wind energy development. In the autumn of 2009, 2010 and 2011, stomach and tissue samples were taken from 20 fish and invertebrate species for analysis of diet composition and δ(15) N and δ(13) C signatures. The food chain in Rhode Island and Block Island Sounds comprises approximately four trophic levels within which the fish community is divided into distinct dietary guilds, including planktivores, benthivores, crustacivores and piscivores. Within these guilds, inter-species isotopic and dietary overlap is high, suggesting that resource partitioning or competitive interactions play a major role in structuring the fish community. Carbon isotopes indicate that most fishes are supported by pelagic phytoplankton, although there is evidence that benthic production also plays a role, particularly for obligate benthivores such as skates Leucoraja spp. This type of analysis is useful for developing an ecosystem-based approach to management, as it identifies species that act as direct links to basal resources as well as species groups that share trophic roles.

A trait-based approach reveals the feeding selectivity of a small endangered Mediterranean fish.

Functional traits are growing in popularity in modern ecology, but feeding studies remain primarily rooted in a taxonomic-based perspective. However, consumers do not have any reason to select their prey using a taxonomic criterion, and prey assemblages are variable in space and time, which makes taxon-based studies assemblage-specific. To illustrate the benefits of the trait-based approach to assessing food choice, we studied the feeding ecology of the endangered freshwater fish Barbus meridionalis. We hypothesized that B. meridionalis is a selective predator which food choice depends on several prey morphological and behavioral traits, and thus, its top-down pressure may lead to changes in the functional composition of in-stream macroinvertebrate communities. Feeding selectivity was inferred by comparing taxonomic and functional composition (13 traits) between ingested and free-living potential prey using the Jacob's electivity index. Our results showed that the fish diet was influenced by 10 of the 13 traits tested. Barbus meridionalis preferred prey with a potential size of 5-10 mm, with a medium-high drift tendency, and that drift during daylight. Potential prey with no body flexibility, conical shape, concealment traits (presence of nets and/or cases, or patterned coloration), and high aggregation tendency had a low predation risk. Similarly, surface swimmers and interstitial taxa were low vulnerable to predation. Feeding selectivity altered the functional composition of the macroinvertebrate communities. Fish absence favored taxa with weak aggregation tendency, weak flexibility, and a relatively large size (10-20 mm of potential size). Besides, predatory invertebrates may increase in fish absence. In conclusion, our study shows that the incorporation of the trait-based approach in diet studies is a promising avenue to improve our mechanistic understanding of predator-prey interactions and to help predict the ecological outcomes of predator invasions and extinctions.

Discovering hidden biodiversity: the use of complementary monitoring of fish diet based on DNA barcoding in freshwater ecosystems.

Ecological monitoring contributes to the understanding of complex ecosystem functions. The diets of fish reflect the surrounding environment and habitats and may, therefore, act as useful integrating indicators of environmental status. It is, however, often difficult to visually identify items in gut contents to species level due to digestion of soft-bodied prey beyond visual recognition, but new tools rendering this possible are now becoming available. We used a molecular approach to determine the species identities of consumed diet items of an introduced generalist feeder, brown trout (Salmo trutta), in 10 Tasmanian lakes and compared the results with those obtained from visual quantification of stomach contents. We obtained 44 unique taxa (OTUs) belonging to five phyla, including seven classes, using the barcode of life approach from cytochrome oxidase I (COI). Compared with visual quantification, DNA analysis showed greater accuracy, yielding a 1.4-fold higher number of OTUs. Rarefaction curve analysis showed saturation of visually inspected taxa, while the curves from the DNA barcode did not saturate. The OTUs with the highest proportions of haplotypes were the families of terrestrial insects Formicidae, Chrysomelidae, and Torbidae and the freshwater Chironomidae. Haplotype occurrence per lake was negatively correlated with lake depth and transparency. Nearly all haplotypes were only found in one fish gut from a single lake. Our results indicate that DNA barcoding of fish diets is a useful and complementary method for discovering hidden biodiversity.

Are parasite richness and abundance linked to prey species richness and individual feeding preferences in fish hosts?

Variations in levels of parasitism among individuals in a population of hosts underpin the importance of parasites as an evolutionary or ecological force. Factors influencing parasite richness (number of parasite species) and load (abundance and biomass) at the individual host level ultimately form the basis of parasite infection patterns. In fish, diet range (number of prey taxa consumed) and prey selectivity (proportion of a particular prey taxon in the diet) have been shown to influence parasite infection levels. However, fish diet is most often characterized at the species or fish population level, thus ignoring variation among conspecific individuals and its potential effects on infection patterns among individuals. Here, we examined parasite infections and stomach contents of New Zealand freshwater fish at the individual level. We tested for potential links between the richness, abundance and biomass of helminth parasites and the diet range and prey selectivity of individual fish hosts. There was no obvious link between individual fish host diet and helminth infection levels. Our results were consistent across multiple fish host and parasite species and contrast with those of earlier studies in which fish diet and parasite infection were linked, hinting at a true disconnect between host diet and measures of parasite infections in our study systems. This absence of relationship between host diet and infection levels may be due to the relatively low richness of freshwater helminth parasites in New Zealand and high host-parasite specificity.