Marine J. Briand

A quick and simple method, usable in the field, for collecting parasites in suitable condition for both morphological and molecular studies

Many methods have been proposed for collecting and fixing parasites, but most were written before the molecular age, and were intended to be practised by experienced parasitologists in well-equipped laboratories. We describe here a very simple method, illustrated by photographs, for collecting helminths from the digestive tract of vertebrates. It only requires a few plastic vials, some ethanol and a means to heat water. Basically, the method consists of: (a) the extraction of all organs from the abdominal cavity; (b) opening the digestive system longitudinally; (c) agitate gut and contents in a saline solution (i.e. ca. 9% NaCl or 1/4 sea water in tap water); (d) decant in saline as many times as needed to clean contents; (e) immediately fix parasites in near-boiling saline; (f) discard saline and keep specimens in 95% ethanol. Additional information is given for collecting parasites from fish gills with a similar process. The method will collect most helminths (digeneans, larval cestodes, nematodes, acanthocephalans) from the digestive tract, and monogeneans and isopod and copepod crustaceans from fish gills. The specimens will be suitable for both morphological study and DNA sequencing. The method is simple, fast, inexpensive and can be used by untrained personnel, even in the field without electricity and without a binocular microscope. It can also be used by trained parasitologists who need to expedite treatment of abundant samples.

Major Sources of Organic Matter in a Complex Coral Reef Lagoon: Identification from Isotopic Signatures (δ13C and δ15N).

A wide investigation was conducted into the main organic matter (OM) sources supporting coral reef trophic networks in the lagoon of New Caledonia. Sampling included different reef locations (fringing, intermediate and barrier reef), different associated ecosystems (mangroves and seagrass beds) and rivers. In total, 30 taxa of macrophytes, plus pools of particulate and sedimentary OM (POM and SOM) were sampled. Isotopic signatures (C and N) of each OM sources was characterized and the composition of OM pools assessed. In addition, spatial and seasonal variations of reef OM sources were examined. Mangroves isotopic signatures were the most C-depleted (-30.17 ± 0.41 ‰) and seagrass signatures were the most C-enriched (-4.36 ± 0.72 ‰). Trichodesmium spp. had the most N-depleted signatures (-0.14 ± 0.03 ‰) whereas mangroves had the most N-enriched signatures (6.47 ± 0.41 ‰). The composition of POM and SOM varied along a coast-to-barrier reef gradient. River POM and marine POM contributed equally to coastal POM, whereas marine POM represented 90% of the POM on barrier reefs, compared to 10% river POM. The relative importance of river POM, marine POM and mangroves to the SOM pool decreased from fringing to barrier reefs. Conversely, the relative importance of seagrass, Trichodesmium spp. and macroalgae increased along this gradient. Overall, spatial fluctuations in POM and SOM were much greater than in primary producers. Seasonal fluctuations were low for all OM sources. Our results demonstrated that a large variety of OM sources sustain coral reefs, varying in their origin, composition and role and suggest that δ13C was a more useful fingerprint than δ15N in this endeavour. This study also suggested substantial OM exchanges and trophic connections between coral reefs and surrounding ecosystems. Finally, the importance of accounting for environmental characteristics at small temporal and spatial scales before drawing general patterns is highlighted.

Anguilliform fish reveal large scale contamination by mine trace elements in the coral reefs of New Caledonia.

Due to intensive mining activity, increasing urbanization and industrialization, vast amounts of contaminants are discharged into the lagoon of New Caledonia, one of the largest continuous coral reef systems and a major biodiversity hotspot. The levels of 11 trace element concentrations were examined in the muscles of predator fish in the south-western lagoon (moray eels and congers). These species are sedentary, widespread, abundant, and they are easily collected using a sea snake sampling technique. We found the highest mean and maximal concentrations of different trace elements ever found in coral fish, notably regarding trace elements typical from mining activity (e.g., mean values for Cr and Ni, respectively: 5.53 ± 6.99 μg g(-1) [max, 35.7 μg g(-1)] and 2.84 ± 3.38 μg g(-1) [max, 18.0 μg g(-1)]). Results show that important trace element contamination extends throughout the lagoon to the barrier reef, following a concentration gradient from the oldest nickel factory (Nouméa).

Variations of natremia in sea kraits (Laticauda spp.) kept in seawater and fresh water.

Marine tetrapods evolved specific excretory structures (e.g. salt glands) that maintain salt concentrations within a narrow range of variation. However, recent investigations showed that in some lineages (sea snakes), individuals dehydrate in seawater and cannot equilibrate their hydromineral balance without access to fresh water. How these marine species cope with salt gain is therefore puzzling. We sampled two species of amphibious sea kraits (Laticauda saintgironsi and L. laticaudata) in the field. We also experimentally investigated patterns of salt regulation, specifically variations in natremia (plasma sodium) and body mass (net water flow), in individuals transferred first to fresh water and then to seawater. Our results show that free-ranging sea kraits display hypernatremia (up to 205mmol·l(-1)). Experimental data showed that natremia markedly decreased in snakes exposed to fresh water and increased when they were transferred to saltwater, thereby demonstrating a marked flexibility in their relation to environmental conditions. A literature survey indicated that all free-ranging marine snake species usually display hypernatremia despite having functional salt glands. Overall, sea snakes exhibit a marked tolerance to salt load compared to other marine tetrapods and apparently trigger substantial salt excretion only once natremia exceeds a high threshold. We hypothesise that this high tolerance significantly decreases energetic costs linked to salt gland functioning.

Do researchers impact their study populations? Assessing the effect of field procedures in a long term population monitoring of sea kraits

Long term population monitoring is essential to ecological studies; however, field procedures may disturb individuals. Assessing this topic is important in worldwide declining taxa such as reptiles. Previous studies focussed on animal welfare issues and examined short-term effects (e.g. increase of stress hormones due to handling). Long-term effects with possible consequences at the population level remain poorly investigated. In the present study, we evaluated the effects of widely used field procedures (e.g. handling, marking, forced regurgitation) both on short-term (hormonal stress response) and on long-term (changes in body condition, survival) scales in two intensively monitored populations of sea kraits (Laticauda spp.) in New Caledonia. Focusing on the most intensively monitored sites, from 2002 to 2012, we gathered approximately 11 200 captures/recaptures on 4500 individuals. Each snake was individually marked (scale clipping + branding) and subjected to various measurements (e.g. body size, head morphology, palpation). In addition, a subsample of more than 500 snakes was forced to regurgitate their prey for dietary analyses. Handling caused a significant stress hormonal response, however we found no detrimental long-term effect on body condition. Forced regurgitation did not cause any significant effect on both body condition one year later and survival. These results suggest that the strong short-term stress provoked by field procedures did not translate into negative effects on the population. Although similar analyses are required to test the validity of our conclusions in other species, our results suggest distinguishing welfare and population issues to evaluate the potential impact of population surveys.

Spatial variability of metallic and organic contamination of anguilliform fish in New Caledonia

New Caledonia is one of the main hot spots of biodiversity on the planet. Large amounts of contaminants are discharged into the lagoon as a result of increasing anthropogenic activities such as intense mining, urbanization, and industrialization. Concentrations of 14 trace elements and 26 persistent organic pollutants (POPs: PCBs and pesticides) were measured in the muscles of two anguilliform fish species, over a coast to barrier reef gradient in two lagoon areas differently exposed to anthropic disturbances. This study emphasizes the high trace element contamination status of anguilliform fish and also highlights slight but perceptible organic pollution. The contamination extends throughout the lagoon, from coast to barrier reef, even in areas remote from emission points. High levels of trace elements, especially those linked to mining activities (i.e., Co, Cr, Fe, Mn, and Ni), were detected in coastal sites. Furthermore, the large dispersion of most POPs throughout the entire lagoon poses the question of their potential toxicity on marine organisms from numerous habitats. Our results underline the need for long-term monitoring of various contaminants over large spatial and time scales.

Occurrence of Anisakis (Nematoda: Anisakidae) larvae in unusual hosts in Southern hemisphere

Nematodes belonging to the genus Anisakis are important parasites due to their abundance in seafood and health impacts on humans. In the present study Anisakis larvae were found in a number of uncommon hosts including the Grey petrel, Procellaria cinerea, the Little penguin, Eudyptula minor, Blue-lipped sea krait, Laticauda laticaudata and Spinner shark, Carcharhinus brevipinna. Morphological examination showed nematodes in these animals are Anisakis larval type I. Genetic characterisation suggested that the larva from one Grey petrel was Anisakis berlandi, whereas the other larvae from the second Grey petrel and from the little penguin were Anisakis pegreffii. A number of larvae found in Blue-lipped sea krait and Spinner shark were identified as Anisakis typica. This is the first report of infective stage of Anisakis larvae parasitising hosts other than teleost fish. Understanding of the extent of infection and the pathogenicity of anisakid nematodes in hosts found in the present study is important in the conservation studies and management plans of these hosts.

Tracking trace elements into complex coral reef trophic networks

The integration, accumulation and transfer of trace elements across the main tropic levels of many food webs are poorly documented. This is notably the case for the complex trophic webs of coral reef ecosystems. Our results show that in the south-west lagoon of New Caledonia both abiotic (i.e. sediments) and biotic (i.e. primary producers, consumers and predators) compartments are contaminated by trace elements. However, our analyses revealed different contamination patterns from the sources of organic matter to the predators. The trophic levels involved in the sedimentary benthic food web (S-BFW, based on the sedimentary organic matter) and to a lesser extent in the reef benthic food web (R-BFW, based on algal turf) were mainly contaminated by trace elements that originate from mining activities like Ni and associated trace elements (Co, Cr, Fe, and Mn). Trace elements linked to agro-industrial (As, Hg, and Zn) and urban (Ag, Cd, Cu, Pb, Se, and V) activities were also integrated into the S-BFW, but preferentially into the R-BFW, and to a lesser extent into the detrital benthic food web (D-BFW, supplied by sea-grass plants). Most of the trace elements were biodiminished with increasing trophic levels along food webs. However, a marked biomagnification was observed for Hg, and suspected for Se and Zn. These results provide important baseline information to better interpret trace element contamination in the different organisms and trophic levels in a highly diversified coral reef lagoon.

Coral assemblages in Tonga: spatial patterns, replenishment capacities, and implications for conservation strategies.

Coral reefs in Tonga have been confronted by multiple threats of various origins, including large-scale disturbances and human-induced stressors. These reef communities have been poorly studied, and efficient conservation actions are urgently needed. The aim of this study was to: (1) examine the spatial distribution of coral assemblages in the lagoon of Tongatapu; (2) determine the degree to which spatial heterogeneity of adult corals is influenced by recruitment processes; and (3) examine the implications of these results in terms of conservation actions. We recorded a total of 37 adult and 28 juvenile coral genera, a mean density of 11.6 adult and 5.5 juvenile colonies m−2, and a dominance of Montipora, Acropora, and Porites. For seven of the 10 dominant genera, spatial patterns of adults were linked to the short-term recruitment pattern history. Despite a reduced diversity and abundance of adult corals in some areas, the lagoon of Tongatapu retains the potential for replenishment through recruitment of young corals. Consequently, we suggest that conservation actions should focus on reducing factors causing coral mortality and maintain suitable conditions for the establishment and growth of juvenile corals, thus increasing the probability that they will reach maturity and participate to the maintenance of local populations. Rather than establishing a large marine protected area, which will almost certainly suffer from a lack of control and poor enforcement, alternative conservation measures could be successfully implemented through the establishment of several small village-based marine reserves, as has been undertaken in other South Pacific islands with promising results.

Catfish Plotosus lineatus are prey for the rare seasnake Hydrophis major in New Caledonia

Seasnakes are potential major predators in coral reef ecosystems (Ineich et al. 2007) but remain poorly studied. In New Caledonia, 14 species of seasnakes are reported but only Laticauda laticaudata and the endemic L. saintgironsi, widely distributed around the island, are abundant on most reefs and islets within the lagoon (Brischoux et al. 2011). Other seasnakes remain largely unstudied, and there are no reliable observations of most species. Hydrophis major is distributed from New Caledonia to South Papua New Guinea and northeast Australia. Observations were made from July 2010 to July 2011, totalizing about 600 h of snorkeling (N 110 dives) at depths of 0–25 m in the SW lagoon of New Caledonia. In total, only 9 H. major were seen, that is, 0.015 individual.h, always between depths of 0 and 5 m (Jack Berthomier, personal communication). The surface area explored can be approximately estimated at 500 m2.h, resulting in a mean density of H. major of 0.3 individual.ha. Thus, each observation of this seasnake is a rare event and must be recorded. Among the 9 seasnakes seen, predation on fish was observed 4 times and always involved Plotosus lineatus. One attack, by a seasnake of about 1.5 m, occurred at dusk and targeted a fish of 25–30 cm TL belonging to a small group of fish (<10 individuals) hiding in a reef cave (Fig. 1a). The other three successful attacks (seasnakes of about 1.0–1.2 m) occurred in the afternoon and focused on several small fish individuals (5 cm TL) belonging to large schools (Fig. 1b). The diet of H. major is poorly known, but the rare existing literature reports P. lineatus as possible prey (Voris and Voris 1983). Our observations, thus, clearly reinforce this view and also highlight that fish venom and spines like those found on Plotosus spp. are not effective defense against predators such as seasnakes.