Coral reef fish density at a tourist destination responded rapidly to COVID-19 restrictions.

Throughout the world, anthropogenic pressure on natural ecosystems is intensifying, notably through urbanisation, economic development, and tourism. Coral reefs have become exposed to stressors related to tourism. To reveal the impact of human activities on fish communities, we used COVID-19-related social restrictions in 2021. In French Polynesia, from February to December 2021, there was a series of restrictions on local activities and international tourism. We assessed the response of fish populations in terms of changes in the species richness and density of fish in the lagoon of Bora-Bora (French Polynesia). We selected sites with varying human pressures-some dedicated to tourism activities, others affected by boat traffic, and control sites with little human presence. Underwater visual surveys demonstrated that fish density and richness differed spatially and temporally. They were lowest on sites affected by boat traffic regardless of pandemic-related restrictions, and when activities were authorised; they were highest during lockdowns. Adult fish density increased threefold on sites usually affected by boat traffic during lockdowns and increased 2.7-fold on eco-tourism sites during international travel bans. Human activities are major drivers of fish density and species richness spatially across the lagoon of Bora-Bora but also temporally across pandemic-related restrictions, with dynamic responses to different restrictions. These results highlight the opportunity provided by pauses in human activities to assess their impact on the environment and confirm the need for sustainable lagoon management in Bora-Bora and similar coral reef settings affected by tourism and boat traffic.

Roadside sales activities in a South Pacific Island (Bora-Bora) reveal sustainable strategies for local food supply during a pandemic.

During the COVID-19 pandemic, the reduced exports and imports as well as the lack of activity due to the interruption in the international tourism economy seriously impacted food security in many Pacific Islands. People often returned to natural resources to provide for themselves, their families, or to generate income. On Bora-Bora Island, the major tourist destination in French Polynesia, roadside sales are widespread. Our study analyses the impact of the COVID-19 pandemic on roadside sales activities through a census of roadside stalls on the five Bora-Bora districts conducted before (January and February 2020), during (from March 2020 to October 2021) and after (from November to December 2021) health-related activity and travel restrictions. Our results showed that the marketing system for local products (fruits, vegetables, cooked meals, and fish) increased in the form of roadside sales during the COVID-19 in two of the five districts of Bora-Bora. Roadside selling would be an alternative system for providing food to the population at Bora-Bora during a global crisis and that could reveal itself sustainable after this pandemic.

Harbours as unique environmental sites of multiple anthropogenic stressors on fish hormonal systems.

Fish development and acclimation to environmental conditions are strongly mediated by the hormonal endocrine system. In environments contaminated by anthropogenic stressors, hormonal pathway alterations can be detrimental for growth, survival, fitness, and at a larger scale for population maintenance. In the context of increasingly contaminated marine environments worldwide, numerous laboratory studies have confirmed the effect of one or a combination of pollutants on fish hormonal systems. However, this has not been confirmed in situ. In this review, we explore the body of knowledge related to the influence of anthropogenic stressors disrupting fish endocrine systems, recent advances (focusing on thyroid hormones and stress hormones such as cortisol), and potential research perspectives. Through this review, we highlight how harbours can be used as "in situ laboratories" given the variety of anthropogenic stressors (such as plastic, chemical, sound, light pollution, and invasive species) that can be simultaneously investigated in harbours over long periods of time.

Long-term exposure to artificial light at night in the wild decreases survival and growth of a coral reef fish.

Artificial light at night (ALAN) is an increasing anthropogenic pollutant, closely associated with human population density, and now well recognized in both terrestrial and aquatic environments. However, we have a relatively poor understanding of the effects of ALAN in the marine realm. Here, we carried out a field experiment in the coral reef lagoon of Moorea, French Polynesia, to investigate the effects of long-term exposure (18-23 months) to chronic light pollution at night on the survival and growth of wild juvenile orange-fin anemonefish, Amphiprion chrysopterus. Long-term exposure to environmentally relevant underwater illuminance (mean: 4.3 lux), reduced survival (mean: 36%) and growth (mean: 44%) of juvenile anemonefish compared to that of juveniles exposed to natural moonlight underwater (mean: 0.03 lux). Our study carried out in an ecologically realistic situation in which the direct effects of artificial lighting on juvenile anemonefish are combined with the indirect consequences of artificial lighting on other species, such as their competitors, predators, and prey, revealed the negative impacts of ALAN on life-history traits. Not only are there immediate impacts of ALAN on mortality, but the decreased growth of surviving individuals may also have considerable fitness consequences later in life. Future studies examining the mechanisms behind these findings are vital to understand how organisms can cope and survive in nature under this globally increasing pollutant.

d-Peptidase Activity in a Marine Mollusk Detoxifies a Nonribosomal Cyclic Lipopeptide: An Ecological Model to Study Antibiotic Resistance.

In the marine environment, sessile cyanobacteria have developed chemical strategies for protection against grazers. In turn, herbivores have to circumvent these defenses and in certain cases even take advantage of them as shelter from their own predators. This is the case of Stylocheilus striatus, a sea hare that feeds on Anabaena torulosa, a cyanobacterium that produces toxic cyclic lipopeptides of the laxaphycin B family. S. striatus consumes the cyanobacterium without being affected by the toxicity of its compounds and also uses it as an invisibility cloak against predators. In this article, using different substrates analogous to laxaphycin B, we demonstrate the presence of an enzyme in the digestive gland of the mollusk that is able to biotransform laxaphycin B derivatives. The enzyme belongs to the poorly known family of d-peptidases that are suspected to be involved in antibiotic resistance.

Hormonal and behavioural effects of motorboat noise on wild coral reef fish.

Anthropogenic noise is an emergent ecological pollutant in both terrestrial and aquatic habitats. Human population growth, urbanisation, resource extraction, transport and motorised recreation lead to elevated noise that affects animal behaviour and physiology, impacting individual fitness. Currently, we have a poor mechanistic understanding of the effects of anthropogenic noise, but a likely candidate is the neuroendocrine system that integrates information about environmental stressors to produce regulatory hormones; glucocorticoids (GCs) and androgens enable rapid individual phenotypic adjustments that can increase survival. Here, we carried out two field-based experiments to investigate the effects of short-term (30 min) and longer-term (48 h) motorboat-noise playback on the behaviour, GCs (cortisol) and androgens of site-attached free-living orange-fin anemonefish (Amphiprion chrysopterus). In the short-term, anemonefish exposed to motorboat-noise playback showed both behavioural and hormonal responses: hiding and aggression increased, and distance moved out of the anemone decreased in both sexes; there were no effects on cortisol levels, but male androgen levels (11-ketotestosterone and testosterone) increased. Some behaviours showed carry-over effects from motorboat noise after it had ceased, and there was no evidence for a short-term change in response to subsequent motorboat-noise playback. Similarly, there was no evidence that longer-term exposure led to changes in response: motorboat noise had an equivalent effect on anemonefish behaviour and hormones after 48 h as on first exposure. Longer-term noise exposure led to higher levels of cortisol in both sexes and higher testosterone levels in males, and stress-responses to an additional environmental challenge in both sexes were impaired. Circulating androgen levels correlated with aggression, while cortisol levels correlated with hiding, demonstrating in a wild population that androgen/glucocorticoid pathways are plausible proximate mechanisms driving behavioural responses to anthropogenic noise. Combining functional and mechanistic studies are crucial for a full understanding of this global pollutant.

Near-future ocean warming and acidification alter foraging behaviour, locomotion, and metabolic rate in a keystone marine mollusc.

Environmentally-induced changes in fitness are mediated by direct effects on physiology and behaviour, which are tightly linked. We investigated how predicted ocean warming (OW) and acidification (OA) affect key ecological behaviours (locomotion speed and foraging success) and metabolic rate of a keystone marine mollusc, the sea hare Stylocheilus striatus, a specialist grazer of the toxic cyanobacterium Lyngbya majuscula. We acclimated sea hares to OW and/or OA across three developmental stages (metamorphic, juvenile, and adult) or as adults only, and compare these to sea hares maintained under current-day conditions. Generally, locomotion speed and time to locate food were reduced ~1.5- to 2-fold when the stressors (OW or OA) were experienced in isolation, but reduced ~3-fold when combined. Decision-making was also severely altered, with correct foraging choice nearly 40% lower under combined stressors. Metabolic rate appeared to acclimate to the stressors in isolation, but was significantly elevated under combined stressors. Overall, sea hares that developed under OW and/or OA exhibited a less severe impact, indicating beneficial phenotypic plasticity. Reduced foraging success coupled with increased metabolic demands may impact fitness in this species and highlight potentially large ecological consequences under unabated OW and OA, namely in regulating toxic cyanobacteria blooms on coral reefs.

Anemone bleaching increases the metabolic demands of symbiont anemonefish.

Increased ocean temperatures are causing mass bleaching of anemones and corals in the tropics worldwide. While such heat-induced loss of algal symbionts (zooxanthellae) directly affects anemones and corals physiologically, this damage may also cascade on to other animal symbionts. Metabolic rate is an integrative physiological trait shown to relate to various aspects of organismal performance, behaviour and locomotor capacity, and also shows plasticity during exposure to acute and chronic stressors. As climate warming is expected to affect the physiology, behaviour and life history of animals, including ectotherms such as fish, we measured if residing in bleached versus unbleached sea anemones (Heteractis magnifica) affected the standard (i.e. baseline) metabolic rate and behaviour (activity) of juvenile orange-fin anemonefish (Amphiprion chrysopterus). Metabolic rate was estimated from rates of oxygen uptake [Formula: see text], and the standard metabolic rate [Formula: see text] of anemonefish from bleached anemones was significantly higher by 8.2% compared with that of fish residing in unbleached anemones, possibly due to increased stress levels. Activity levels did not differ between fish from bleached and unbleached anemones. As [Formula: see text] reflects the minimum cost of living, the increased metabolic demands may contribute to the negative impacts of bleaching on important anemonefish life history and fitness traits observed previously (e.g. reduced spawning frequency and lower fecundity).

Ephemeral and Localized Outbreaks of the Coral Predator Acanthaster cf. solaris in the Southwestern Lagoon of New Caledonia.

Mehdi Adjeroud, Mohsen Kayal, Christophe Peignon, Matthieu Juncker, Suzanne C. Mills, Ricardo Beldade, and Pascal Dumas (2018) Outbreaks of the coral predator Acanthaster spp., the crown-of-thorns seastar (COTS), cause major coral declines across the Indo-Pacific. However, the processes surrounding the initiation and propagation of COTS outbreaks are still unclear. We observed COTS outbreak abundances on several mid-shelf and inner-barrier reefs in the southern section of the New Caledonian lagoon, a multi-location initiation event that we expected to precede a broader region-wide disturbance. However, reef monitoring over 3 years revealed the highly localized and ephemeral character of these outbreaks. Outbreaks that were observed at four reef locations at the beginning of the survey simply faded away, without any specific management actions such as culling efforts. We also found no distinct reef biotope on which COTS outbreaks originated, although mid-shelf and inner-shelf barrier reefs seem to be favoured. New Caledonia has been exempted from to widespread regional COTS outbreaks, which is surprising given its proximity to the Australian Great Barrier Reef - a reef complex particularly vulnerable to COTS - and Vanuatu - where large-scale outbreaks were recorded during the same period. The availability of coral prey is probably not a limiting factor for the propagation of COTS outbreaks on New Caledonian reefs, given the high abundance and diversity of coral assemblages. Our findings reveal that localized and ephemeral COTS outbreaks can be naturally contained and do not necessarily result in widespread disturbances.

Cascading effects of thermally-induced anemone bleaching on associated anemonefish hormonal stress response and reproduction.

Organisms can behaviorally, physiologically, and morphologically adjust to environmental variation via integrative hormonal mechanisms, ultimately allowing animals to cope with environmental change. The stress response to environmental and social changes commonly promotes survival at the expense of reproduction. However, despite climate change impacts on population declines and diversity loss, few studies have attributed hormonal stress responses, or their regulatory effects, to climate change in the wild. Here, we report hormonal and fitness responses of individual wild fish to a recent large-scale sea warming event that caused widespread bleaching on coral reefs. This 14-month monitoring study shows a strong correlation between anemone bleaching (zooxanthellae loss), anemonefish stress response, and reproductive hormones that decreased fecundity by 73%. These findings suggest that hormone stress responses play a crucial role in changes to population demography following climate change and plasticity in hormonal responsiveness may be a key mechanism enabling individual acclimation to climate change.Elevated temperatures can cause anemones to bleach, with unknown effects on their associated symbiotic fish. Here, Beldade and colleagues show that climate-induced bleaching alters anemonefish hormonal stress response, resulting in decreased reproductive hormones and severely impacted reproduction.

Motorboat noise disrupts co-operative interspecific interactions.

Human-made noise is contributing increasingly to ocean soundscapes. Its physical, physiological and behavioural effects on marine organisms are potentially widespread, but our understanding remains largely limited to intraspecific impacts. Here, we examine how motorboats affect an interspecific cleaning mutualism critical for coral reef fish health, abundance and diversity. We conducted in situ observations of cleaning interactions between bluestreak cleaner wrasses (Labroides dimidiatus) and their fish clients before, during and after repeated, standardised approaches with motorboats. Cleaners inspected clients for longer and were significantly less cooperative during exposure to boat noise, and while motorboat disturbance appeared to have little effect on client behaviour, as evidenced by consistency of visit rates, clientele composition, and use of cleaning incitation signals, clients did not retaliate as expected (i.e., by chasing) in response to increased cheating by cleaners. Our results are consistent with the idea of cognitive impairments due to distraction by both parties. Alternatively, cleaners might be taking advantage of distracted clients to reduce their service quality. To more fully understand the importance of these findings for conservation and management, further studies should elucidate whether the efficacy of ectoparasite removal by cleaners is affected and explore the potential for habituation to boat noise in busy areas.

The embryonic life history of the tropical sea hare Stylocheilus striatus (Gastropoda: Opisthobranchia) under ambient and elevated ocean temperatures.

Ocean warming represents a major threat to marine biota worldwide, and forecasting ecological ramifications is a high priority as atmospheric carbon dioxide (CO2) emissions continue to rise. Fitness of marine species relies critically on early developmental and reproductive stages, but their sensitivity to environmental stressors may be a bottleneck in future warming oceans. The present study focuses on the tropical sea hare, Stylocheilus striatus (Gastropoda: Opisthobranchia), a common species found throughout the Indo-West Pacific and Atlantic Oceans. Its ecological importance is well-established, particularly as a specialist grazer of the toxic cyanobacterium, Lyngbya majuscula. Although many aspects of its biology and ecology are well-known, description of its early developmental stages is lacking. First, a detailed account of this species' life history is described, including reproductive behavior, egg mass characteristics and embryonic development phases. Key developmental features are then compared between embryos developed in present-day (ambient) and predicted end-of-century elevated ocean temperatures (+3 °C). Results showed developmental stages of embryos reared at ambient temperature were typical of other opisthobranch species, with hatching of planktotrophic veligers occurring 4.5 days post-oviposition. However, development times significantly decreased under elevated temperature, with key embryonic features such as the velum, statocysts, operculum, eyespots and protoconch developing approximately 24 h earlier when compared to ambient temperature. Although veligers hatched one day earlier under elevated temperature, their shell size decreased by approximately 20%. Our findings highlight how an elevated thermal environment accelerates planktotrophic development of this important benthic invertebrate, possibly at the cost of reducing fitness and increasing mortality.

Evolutionary Conflict Between Maternal and Paternal Interests: Integration with Evolutionary Endocrinology.

Conflict between mates, as well as conflict between parents and offspring are due to divergent evolutionary interests of the interacting individuals. Hormone systems provide genetically based proximate mechanisms for mediating phenotypic adaptation and maladaptation characteristic of evolutionary conflict between individuals. Testosterone (T) is among the most commonly studied hormones in evolutionary biology, and as such, its role in shaping sexually dimorphic behaviors and physiology is relatively well understood, but its role in evolutionary conflict is not as clear. In this review, we outline the genomic conflicts arising within the family unit, and incorporate multiple lines of evidence from the bank vole (Myodes glareolus) system to outline how T impacts traits associated with reproduction and survival, resulting in a sexually antagonistic genetic trade-off in fitness. A major prediction arising from this work is that lower T is favored in females, whereas the optimal T level in males fluctuates in relation to social and ecological factors. We additionally discuss future directions to further integrate endocrinology into the study of sexual and parent-offspring conflicts.

Repeated exposure to noise increases tolerance in a coral reef fish.

Some anthropogenic noise is now considered pollution, with evidence building that noise from human activities such as transportation, construction and exploration can impact behaviour and physiology in a broad range of taxa. However, relatively little research has considered the effects of repeated or chronic noise; extended exposures may result in habituation or sensitisation, and thus changes in response. We conducted a field-based experiment at Moorea Island to investigate how repeated exposure to playback of motorboat noise affected a coral reef fish (Dascyllus trimaculatus). We found that juvenile D. trimaculatus increased hiding behaviour during motorboat noise after two days of repeated exposure, but no longer did so after one and two weeks of exposure. We also found that naïve individuals responded to playback of motorboat noise with elevated ventilation rates, but that this response was diminished after one and two weeks of repeated exposure. We found no strong evidence that baseline blood cortisol levels, growth or body condition were affected by three weeks of repeated motorboat-noise playback. Our study reveals the importance of considering how tolerance levels may change over time, rather than simply extrapolating from results of short-term studies, if we are to make decisions about regulation and mitigation.

Isolation and Synthesis of Laxaphycin B-Type Peptides: A Case Study and Clues to Their Biosynthesis.

The laxaphyci's B family constitutes a group of five related cyclic lipopeptides isolated from diverse cyanobacteria from all around the world. This group shares a typical structure of 12 amino acids from the l and d series, some of them hydroxylated at the beta position, and all containing a rare beta-amino decanoic acid. Nevertheless, they can be differentiated due to slight variations in the composition of their amino acids, but the configuration of their alpha carbon remains conserved. Here, we provide the synthesis and characterization of new laxaphycin B-type peptides. In doing so we discuss how the synthesis of laxaphycin B and analogues was developed. We also isolate minor acyclic laxaphycins B, which are considered clues to their biosynthesis.

Lime Juice and Vinegar Injections as a Cheap and Natural Alternative to Control COTS Outbreaks.

Outbreaks of the corallivorous crown-of-thorns seastar Acanthaster planci (COTS) represent one of the greatest disturbances to coral reef ecosystems in the Indo-Pacific, affecting not only coral reefs but also the coastal communities which rely on their resources. While injection approaches are increasingly used in an attempt to control COTS densities, most of them display severe drawbacks including logistical challenges, high residual environmental impacts or low cost-effectiveness. We tested a new alternative control method based upon acidic injections of cheap, 100% natural products. We investigated the lethal doses, intra- and inter-specific disease transmission and immune responses of COTS when injected with fresh lime juice (extracted from local Citrus arantifolia) and white spirit vinegar. High COTS mortality was achieved with small volumes: 10-20 ml per seastar induced death in 89%/97% of injected specimens after an average 34.3 h/29.8 h for lime juice and vinegar respectively. Highest efficiency was reached for both solutions with double shots of (2 × 10 ml) in two different areas on the body: 100% mortality occurred within 12-24 h, which is similar or faster compared with other current injection methods. Multiple immune measures suggested that death was very likely caused by pH stress from the acidic solutions rather than a bacterial infection. Contagion to either conspecifics or a variety of other reef species was not observed, even at COTS densities 15 times higher than the highest naturally reported. 10 to 20 l lime juice/vinegar could kill up to a thousand COTS at a cost of less than 0.05 USD per specimen; no permits or special handling procedures are required. We conclude that injections of lime juice and vinegar offer great advantages when compared to current best practises and constitute a cheap and natural option for all reefs affected by COTS.

Metabarcoding dietary analysis of coral dwelling predatory fish demonstrates the minor contribution of coral mutualists to their highly partitioned, generalist diet.

Understanding the role of predators in food webs can be challenging in highly diverse predator/prey systems composed of small cryptic species. DNA based dietary analysis can supplement predator removal experiments and provide high resolution for prey identification. Here we use a metabarcoding approach to provide initial insights into the diet and functional role of coral-dwelling predatory fish feeding on small invertebrates. Fish were collected in Moorea (French Polynesia) where the BIOCODE project has generated DNA barcodes for numerous coral associated invertebrate species. Pyrosequencing data revealed a total of 292 Operational Taxonomic Units (OTU) in the gut contents of the arc-eye hawkfish (Paracirrhites arcatus), the flame hawkfish (Neocirrhites armatus) and the coral croucher (Caracanthus maculatus). One hundred forty-nine (51%) of them had species-level matches in reference libraries (>98% similarity) while 76 additional OTUs (26%) could be identified to higher taxonomic levels. Decapods that have a mutualistic relationship with Pocillopora and are typically dominant among coral branches, represent a minor contribution of the predators' diets. Instead, predators mainly consumed transient species including pelagic taxa such as copepods, chaetognaths and siphonophores suggesting non random feeding behavior. We also identified prey species known to have direct negative interactions with stony corals, such as Hapalocarcinus sp, a gall crab considered a coral parasite, as well as species of vermetid snails known for their deleterious effects on coral growth. Pocillopora DNA accounted for 20.8% and 20.1% of total number of sequences in the guts of the flame hawkfish and coral croucher but it was not detected in the guts of the arc-eye hawkfish. Comparison of diets among the three fishes demonstrates remarkable partitioning with nearly 80% of prey items consumed by only one predator. Overall, the taxonomic resolution provided by the metabarcoding approach highlights a highly complex interaction web and demonstrates that levels of trophic partitioning among coral reef fishes have likely been underestimated. Therefore, we strongly encourage further empirical approaches to dietary studies prior to making assumptions of trophic equivalency in food web reconstruction.

More coral, more fish? Contrasting snapshots from a remote Pacific atoll.

Coral reefs are in decline across the globe as a result of overexploitation, pollution, disease and, more recently, climate change. The impacts of changes in coral cover on associated fish communities can be difficult to predict because of the uneven dependence of reef fish species on corals for food, shelter or the three-dimensional structure they provide. We compared live coral cover, reef fish community metrics, and their associations in two surveys of the lagoon of the remote atoll of Mataiva (French Polynesia) carried out 31 years apart. In contrast to the general pattern of decreasing coral cover reported for many parts of the Indo-Pacific region, live coral cover increased 6-7 fold at Mataiva between 1981 and 2012, and fish density nearly doubled. The stable overall reef fish species richness belied a significant shift in community structure. There was little overlap in community composition across years, and fish assemblages in 2012 were more homogeneous in composition than they were in 1981. Changes in species abundance were not clearly related to species-specific reliance on corals. The strong positive relationships between live coral cover and fish diversity and abundance noted in 1981, when coral cover rarely exceeded 10%, were no longer present in 2012, when coral cover rarely fell below this value. The most parsimonious explanation for these contrasting relationships is that, over the combined range of coral cover observed in the 1981 and 2012 snapshots, there is a rapidly asymptotic relationship between coral and fish. Our results, and other data from the south and west Pacific, suggest that fish diversity and abundance might accumulate rapidly up to a threshold of approximately 10% live coral cover. Such a relationship would have implications for our expectations of resistance and recovery of reef fish communities facing an increasingly severe regime of coral reef disturbances.

Anthropogenic noise playback impairs embryonic development and increases mortality in a marine invertebrate.

Human activities can create noise pollution and there is increasing international concern about how this may impact wildlife. There is evidence that anthropogenic noise may have detrimental effects on behaviour and physiology in many species but there are few examples of experiments showing how fitness may be directly affected. Here we use a split-brood, counterbalanced, field experiment to investigate the effect of repeated boat-noise playback during early life on the development and survival of a marine invertebrate, the sea hare Stylocheilus striatus at Moorea Island (French Polynesia). We found that exposure to boat-noise playback, compared to ambient-noise playback, reduced successful development of embryos by 21% and additionally increased mortality of recently hatched larvae by 22%. Our work, on an understudied but ecologically and socio-economically important taxon, demonstrates that anthropogenic noise can affect individual fitness. Fitness costs early in life have a fundamental influence on population dynamics and resilience, with potential implications for community structure and function.

A new versatile primer set targeting a short fragment of the mitochondrial COI region for metabarcoding metazoan diversity: application for characterizing coral reef fish gut contents.

INTRODUCTION: The PCR-based analysis of homologous genes has become one of the most powerful approaches for species detection and identification, particularly with the recent availability of Next Generation Sequencing platforms (NGS) making it possible to identify species composition from a broad range of environmental samples. Identifying species from these samples relies on the ability to match sequences with reference barcodes for taxonomic identification. Unfortunately, most studies of environmental samples have targeted ribosomal markers, despite the fact that the mitochondrial Cytochrome c Oxidase subunit I gene (COI) is by far the most widely available sequence region in public reference libraries. This is largely because the available versatile ("universal") COI primers target the 658 barcoding region, whose size is considered too large for many NGS applications. Moreover, traditional barcoding primers are known to be poorly conserved across some taxonomic groups. RESULTS: We first design a new PCR primer within the highly variable mitochondrial COI region, the "mlCOIintF" primer. We then show that this newly designed forward primer combined with the "jgHCO2198" reverse primer to target a 313 bp fragment performs well across metazoan diversity, with higher success rates than versatile primer sets traditionally used for DNA barcoding (i.e. LCO1490/HCO2198). Finally, we demonstrate how the shorter COI fragment coupled with an efficient bioinformatics pipeline can be used to characterize species diversity from environmental samples by pyrosequencing. We examine the gut contents of three species of planktivorous and benthivorous coral reef fish (family: Apogonidae and Holocentridae). After the removal of dubious COI sequences, we obtained a total of 334 prey Operational Taxonomic Units (OTUs) belonging to 14 phyla from 16 fish guts. Of these, 52.5% matched a reference barcode (>98% sequence similarity) and an additional 32% could be assigned to a higher taxonomic level using Bayesian assignment. CONCLUSIONS: The molecular analysis of gut contents targeting the 313 COI fragment using the newly designed mlCOIintF primer in combination with the jgHCO2198 primer offers enormous promise for metazoan metabarcoding studies. We believe that this primer set will be a valuable asset for a range of applications from large-scale biodiversity assessments to food web studies.