Kelp detritus: Unutilized productivity or an unacknowledged trophic resource?

Kelp beds are one of the most productive marine systems and, while little of this production is directly consumed, there is growing evidence that kelp detritus is an essential food source for many detrital and suspension feeders, and forms an important component of offshore sedimentary carbon pools. However, the extent of the contribution of kelp detritus to the nutrition of coastal fauna is not well resolved. In this study, we compare the contribution of phytoplankton, kelp detritus, and waste from fish cages to the diet of a sentinel suspension feeder, the blue mussel (Mytilus edulis) using stable isotopes. We found a significant depletion in both 13C and 15N in kelp tissue with age (distance from stipe to the deteriorating distal end of the kelp frond) which may have biased dietary estimates in previous studies which have applied isotopic source values derived from fresh kelp. Our mixing models indicate that macroalgal detritus formed 59% of the diet of the mussels in Berehaven, Bantry Bay, Ireland. We support the isotopic mixing model results by modelling the relative production of phytoplankton, kelp, and salmon farm waste, and found the supply of C and N from kelp and phytoplankton far exceeded the requirements of the mussels with much less coming from the nearby fish cages. Monthly chlorophyll measurements indicated there was only sufficient phytoplankton density to support mussel growth during the spring and autumn, explaining our observation of patterns in the relative importance of utilization of kelp detritus. Where there is pressure to harvest kelp beds, this study highlights the supporting ecosystem service they provide as an important dietary source in coastal food webs and emphasises the need for appropriate management measures for this resource.

Effectiveness of cooking to reduce norovirus and infectious F-specific RNA bacteriophage concentrations in Mytilus edulis.

AIMS: The aim of this study was to determine if domestic cooking practices can reduce concentrations of norovirus (NoV) and F-specific RNA (FRNA) bacteriophage in experimentally contaminated mussels. METHODS AND RESULTS: Mussels (n = 600) contaminated with NoV and FRNA bacteriophage underwent four different cooking experiments performed in triplicate at ~70°C and >90°C. Concentrations of infectious FRNA bacteriophage (using a plaque assay) were compared with concentrations of FRNA bacteriophage and NoV determined using a standardised RT-qPCR. Initial concentrations of infectious FRNA bacteriophage (7·05 log10  PFU g(-1) ) in mussels were not significantly reduced in simmering water (~70°C); however, cooking at higher temperatures (>90°C) reduced infectious FRNA bacteriophage to undetected levels within 3 min. Further investigation determined the time required for a 1-log reduction of infectious FRNA bacteriophage at 90°C to be 42 s therefore a >3-log reduction in infectious virus can be obtained by heating mussel digestive tissue to 90°C for 126 s. CONCLUSIONS: Domestic cooking practices based on shell opening alone do not inactivate infectious virus in mussels, however, cooking mussels at high temperatures is effective to reduce infectious virus concentrations and the risk of illness in consumers. SIGNIFICANCE AND IMPACT OF THE STUDY: The data will contribute towards evidence-based cooking recommendations for shellfish to provide a safe product for human consumption.

The health status of mussels, Mytilus spp., in Ireland and Wales with the molecular identification of a previously undescribed haplosporidian.

Both wild and cultured mussels (Mytilus edulis, Mytilus galloprovincialis and hybrids), are found along most of the Irish coastline. M. edulis is widespread along all Irish coasts and is the only mussel species present on both the east coast of Ireland and the Welsh coast in the Irish Sea. M. galloprovincialis and hybrids are found along the Irish coastline except for the east coast. Samples of Mytilus spp. were collected from twenty-four sites, encompassing all coasts of Ireland and the Welsh coast, at different times of the year and over several years (2008-2011). In total, 841 mussels were examined histologically to assess their health status and the presence of any parasites or commensals. Mussels from 14 of the 24 sites were screened using polymerase chain reaction (PCR) to determine which mytilid species were present. A range of parasites were observed, generally at low levels. The most diverse community of parasites was observed at a sheltered site with poor water quality. Of significance, a previously undescribed haplosporidian was detected in a single mussel sample in the Menai Strait, Wales, by PCR and was confirmed by direct sequencing and is most closely related to Minchina chitonis and a haplosporidian of the Florida marsh clam Cyrenoida floridana. While M. edulis were infected by a variety of micro- and macro-parasites, only trematodes were observed in M. galloprovincialis and hybrids. Habitat description and the environmental factors influencing the study sites, including water quality and exposure, were recorded.

Grazing intensity is a poor indicator of waterborne Escherichia coli O157 activity.

Contamination of watercourses with fecal matter represents a significant risk to public health due to the associated risk from human pathogens (e.g. Escherichia coli O157, norovirus). In addition, water contamination may also perpetuate the re-infection cycle of human pathogens within domesticated and wild animal populations. While diffuse pollution from agricultural fields has been identified as a major source of these pathogens, the relationship between livestock grazing intensity and subsequent pathogen persistence in water is not well established. Our aim was to critically evaluate the importance of land use management on the activity of E. coli O157 in freshwaters collected from a livestock dominated catchment in the UK. We inoculated replicate batches of both filter-sterilised and non-sterile freshwaters with a chromosomally lux-marked E. coli O157 and monitored pathogen survival and activity over a 5 d period. Our results indicate that the greatest risk for pathogens entering freshwater is probably associated with high intensity livestock areas, although their subsequent survival is greatest in waters from low intensity livestock areas. We ascribe this enhanced persistence in the latter to reduced competition and predation within these aquatic environments. These results have serious implications for the reliability of pathogen risk exposure maps which are based on grazing intensity alone.

Alberta Stroke Program Early CT Scoring of CT perfusion in early stroke visualization and assessment.

BACKGROUND AND PURPOSE: Qualitative CT perfusion (CTP) assessment by using the Alberta Stroke Program Early CT Score (ASPECTS) allows rapid calculation of infarct extent for middle cerebral artery infarcts. Published thresholds exist for noncontrast CT (NCCT) ASPECTS, which may distinguish outcome/complication risk, but early ischemic signs are difficult to detect. We hypothesized that different ASPECTS thresholds exist for CTP parameters versus NCCT and that these may be superior at predicting clinical and radiologic outcome in the acute setting. MATERIALS AND METHODS: Thirty-six baseline acute stroke NCCT and CTP studies within 3 hours of symptoms were blindly reviewed by 3 neuroradiologists, and ASPECTS were assigned. Treatment response was defined as major neurologic improvement when a > or =8-point National Institutes of Health Stroke Scale improvement at 24 hours occurred. Follow-up NCCT ASPECTS and 90-day modified Rankin score (mRS) were radiologic and clinical reference standards. Receiver operating characteristic curves derived optimal thresholds for outcome. RESULTS: Cerebral blood volume and NCCT ASPECTS had similar radiologic correlations (0.6 and 0.5, respectively) and best predicted infarct size in the absence of major neurologic improvement. A NCCT ASPECT threshold of 7 and a cerebral blood volume threshold of 8 discriminated patients with poor follow-up scans (P < .0002 and P = .0001) and mRS < or =2 (P = .001 and P < .001). Only cerebral blood volume predicted major neurologic improvement (P = .02). Interobserver agreement was substantial (intraclass correlation coefficient, 0.69). Cerebral blood volume ASPECTS sensitivity, specificity, positive predictive value, and negative predictive value for clinical outcome were 60%, 100%, 100%, and 45%, respectively. No patients with cerebral blood volume ASPECTS <8 achieved good clinical outcome. CONCLUSION: Cerebral blood volume ASPECTS is equivalent to NCCT for predicting radiologic outcome but may have an additional benefit in predicting patients with major neurologic improvement.

A Vibrio splendidus strain is associated with summer mortality of juvenile oysters Crassostrea gigas in the Bay of Morlaix (North Brittany, France).

Juvenile oysters Crassostrea gigas cultured in the Bay of Morlaix (France) have suffered unexplained summer mortalities for over a decade. In the present study, we tested the hypothesis that a bacterial pathogen could be responsible for this phenomenon. A first attempt failed to isolate a bacterial pathogen from moribund or weak oysters. Only non-pathogenic, probably opportunistic, bacteria were isolated. As an alternative approach, we focused on oysters presenting reduced stress-response capacities (determined by circulating noradrenaline measurements), a characteristic of juvenile oysters entering an early phase of the disease. Cultures of bacterial isolates on TCBS plates revealed that a Vibrio strain was present in diseased oysters and scarce or absent in healthy oysters. Experimental infections indicated that this Vibrio can cause mortalities of juvenile oysters when injected at concentrations ranging from 10(4) to 10(8) CFU oyster(-1). Similarly to the summer mortality disease, the Vibrio isolate caused higher mortalities at higher temperatures; apparently, it could not be transmitted horizontally, it did not affect adult oysters and it induced stress-response dysfunctions in juvenile oysters. Phenotypic and genotypic characterizations identified the pathogen as Vibrio splendidus. Taken together, the present results satisfy Koch's postulate and suggest that this bacterial strain is probably responsible for the juvenile oyster summer mortalities in the Bay of Morlaix.

Noradrenaline modulates oyster hemocyte phagocytosis via a beta-adrenergic receptor-cAMP signaling pathway.

Catecholamines (CA) regulate several physiological processes in molluscs. Experiments have been conducted to determine the effects of noradrenaline (NA), the principal CA circulating in bivalve hemolymph, on oyster hemocytephagocytosis. Results show that NA had a dose-dependent inhibitory effect on phagocytosis at physiological concentrations of 0.1 microM and above. The beta-adrenoceptor agonist isoproterenol mimicked the inhibitory effects of NA on phagocytosis, whereas the alpha-adrenoceptor agonist phenylephrine had no significant effect. Furthermore, the beta-adrenoceptor antagonist propanolol, but not the alpha-adrenoceptor antagonist prazosin, prevented the inhibition of phagocytosis by NA. The type IV phosphodiesterase inhibitor rolipram acted synergistically with a suboptimal concentration of isoproterenol to inhibit phagocytosis, and the protein kinase A inhibitor H-89, but not the protein kinase C inhibitor calphostin C, attenuated the effect of isoproterenol. These results show that NA can modulate oyster hemocyte phagocytosis via a beta-adrenergic receptor/cAMP/protein kinase A signaling pathway.

Stress-induced catecholamine changes in the hemolymph of the oyster Crassostrea gigas.

The stress response is a series of coordinated physiological reactions increasing an organism's capacity to maintain homeostasis in the presence of threatening agents. This fundamental process is known to involve hormonal signaling to rapidly modulate key physiological functions in vertebrates, but data are lacking concerning neuroendocrine responses to stress in invertebrates. The present study examined circulating catecholamine (CA) responses to stress in oysters. Mechanical disturbances (consisting of shaking the animals) and temperature or salinity variations were applied to the animals because these three types of stressors are commonly encountered by oysters in aquaculture or in their natural habitat. Results show that both circulating noradrenaline (NA) and dopamine (DA) concentrations increased in response to stress. The catecholaminergic response to acute mechanical stressors was rapid (less than 5 min), transient (a return to basal CA levels was observed after 60-90 min), and reflected both the intensity and duration of the perturbation. In contrast, responses to temperature and salinity variations were long lasting (up to 72 h). CA concentrations varied from 1.61 +/- 0.30 ng NA/ml and 0.41 +/- 0.05 ng DA/ml to maximal values of 22.07 +/- 0.97 ng NA/ml and 2.24 +/- 0.19 ng DA/ml. Such CA concentrations are known to induce physiological responses in bivalves, suggesting that stress-induced NA and DA changes exert a regulatory function in oysters.

Stress and stress-induced neuroendocrine changes increase the susceptibility of juvenile oysters (Crassostrea gigas) to Vibrio splendidus.

Oysters are permanently exposed to various microbes, and their defense system is continuously solicited to prevent accumulation of invading and pathogenic organisms. Therefore, impairment of the animal's defense system usually results in mass mortalities in cultured oyster stocks or increased bacterial loads in food products intended for human consumption. In the present study, experiments were conducted to examine the effects of stress on the juvenile oyster's resistance to the oyster pathogen Vibrio splendidus. Oysters (Crassostrea gigas) were challenged with a low dose of a pathogenic V. splendidus strain and subjected to a mechanical stress 3 days later. Both mortality and V. splendidus loads increased in stressed oysters, whereas they remained low in unstressed animals. Injection of noradrenaline or adrenocorticotropic hormone, two key components of the oyster neuroendocrine stress response system, also caused higher mortality and increased accumulation of V. splendidus in challenged oysters. These results suggest that the physiological changes imposed by stress, or stress hormones, influenced host-pathogen interactions in oysters and increased juvenile C. gigas vulnerability to Vibrio splendidus.

Evidence for a form of adrenergic response to stress in the mollusc Crassostrea gigas.

Catecholamines and pro-opiomelanocortin (POMC)-derived peptides, some of the central regulators of the stress-response systems of vertebrates, are also present in invertebrates. However, studies are needed to determine how these hormones participate in the organisation of neuroendocrine stress-response axes in invertebrates. Our present work provides evidence for the presence of an adrenergic stress-response system in the oyster Crassostrea gigas. Noradrenaline and dopamine are released into the circulation in response to stress. Storage and release of these hormones take place in neurosecretory cells presenting morphological and biochemical similarities with vertebrate chromaffin cells. Both in vivo and in vitro experiments showed that applications of the neurotransmitters acetylcholine or carbachol caused no significant release of noradrenaline or dopamine. Moreover, the nicotinic antagonists hexamethonium and &agr; -bungarotoxin and the muscarinic antagonist atropine caused no significant inhibition of catecholamine release in stressed oysters. Adrenocorticotropic hormone (ACTH) induced a significant release of noradrenaline, but the release of dopamine in response to ACTH was not significant. These results suggest that, unlike that of vertebrates, the adrenergic stress-response system of oysters is not under the control of acetylcholine and that other factors, such as the neuropeptide ACTH, might control this system.

Noradrenaline modulates hemocyte reactive oxygen species production via beta-adrenergic receptors in the oyster Crassostrea gigas.

Catecholamines (CA) are known to be present in the microenvironment of molluscan immunocytes. In the present study, experiments were conducted to determine the effects of noradrenaline (NA), the principal CA circulating in bivalve hemolymph, on the luminol-dependent chemiluminescence (CL) of oyster Crassostrea gigas hemocytes. Results show that NA had a dose-dependent inhibitory effect on the CL-response at the physiological concentration of 0.1 microM and above. The alpha-adrenoceptor agonist phenylephrine had no significant effect on the CL-response whereas the beta-adrenoceptor agonist isoproterenol mimicked the inhibitory effects of NA on the CL-response. The beta-adrenoceptor antagonist propanolol, but not the alpha-adrenoceptor antagonist prazosin, prevented the negative effects of NA on the CL-response. Taken together, these results show that beta-adrenergic receptors are present at the surface of oyster hemocytes and allow NA to down-regulate the CL-response.

Lysozyme and antiprotease activity in the lesser octopus Eledone cirrhosa (Lam.) (Cephalopoda).

Antiprotease and lysozyme activities were detected in various tissue samples including the haemocytes and haemolymph of Eledone cirrhosa. Injection of live Vibrio anguillarum caused an increase in lysozyme activity in the branchial heart over 48 hours and a decrease in the lysozyme activity of haemocytes over 24 hours. Haemocytes from control PBS injected animals demonstrated increased lysozyme levels 4 hours after injection whereas it decreased after the injection of live bacteria in PBS. The lysozyme activity of the haemolymph was not affected by these procedures. Bacteria injections had no effect on the antiprotease activity of the organ samples but increased the antiprotease activity of the haemocytes compared to controls in the 4 h samples. Haemolymph antiprotease activity decreased at a greater rate following bacteria injection than in control PBS injected animals. Haemocyte numbers/ml increased for both the control and bacteria injected animals with a greater increase demonstrated for the bacteria injected animals in the 4 h sample. Concomittant with the increase in the numbers of circulating haemocytes live V. anguillarum were cleared from the circulation of E. cirrhosa in less than 4 hours.