Aspects of the feeding behaviour of intensively reared trout

Feeding behaviour of trained rainbow trout was investigated by the use of demand feeders, under different light conditions. The effects of the energy content of diet, and the size, colour and texture of feed pellets, on the feeding behaviour, were studied. An attempt was made to locate the assumed centres for feeding and satiety in the hypothalamus of brain by the intraperitoneal injections of goldthioglucose. Feeding under nine different constant photoperiods at 160 lux, at a temperature of 13.5°C, showed that trout exhibit a rhythmic pattern of feeding behaviour in all photoperiods except in continuous darkness.Feeding rhythms of trout attributable to the degree of gut distension were formed every eight to ten hours. Further studies by varying levels of light intensity revealed the interaction of light intensity and photoperiod. At shorter photoperiods lower levels of light intensity decreased the feeding activity in terms of food intake but by increasing the photoperiod the same feeding activity was accomplished as by the fish subject to a short photoperiod but under higher light intensity.Simulated effect of increasing and decreasing daylengths did not affect the overall food intake and growth performance. Trout are quite efficient in adjusting their food intake in terms of energy content. Colour, size and texture of feed pellets affect the feeding responses and elicit preferential food selection behaviour in trout. Goldthioglucose induced some reversable toxic effects upon general physiology of trout and did not produce any lesions in the assumed areas of feeding and satiety centres in the brain.  It was concluded that the feeding behaviour of trout exhibited selective preferences according to the physical nature of food items and those preferences could be further influenced by the biotic and abiotic factors, light being one of the most important abiotic factors.

Bacterial associations with salmonid eggs

Rainbow trout eggs Salmo gairdneri, Richardson, were incubated under a range of different environmental conditions. Recovery of bacteria from egg surfaces revealed that increased water temperature, slow water flow rates and high egg density all significantly increased egg surface bacterial populations. Live eggs were mainly colonized by Cytophaga sp., pseudomonas fluorescens and Aeromonas hydrophila. In contrast, dead eggs supported considerable numbers of fluorescent Pseudomonas sp. Analysis of potential nutrient sources for bacteria colonizing live egg surfaces revealed that small amounts of amino acids, phosphate and potassium may be lost by incubating eggs. Subsequently these nutrients were shown to be capable of supporting limited bacterial growth and reproduction. Dead eggs `leaked' increased amounts of the above nutrients which in turn supported higher bacterial numbers. In addition, biochemical analysis of eggs revealed amino acids and fatty acids that might be utilized by bacteria colonizing dead egg surfaces. Assessment of adhesion properties of bacteria frequently recovered from egg surfaces revealed high cell surface hydrophobicity as an important factor in successful egg colonization. Analysis of egg mortalities from groups of rainbow trout and brown trout (S.trutta L.) eggs maintained under two different incubation systems revealed that potentially a close correlation existed between egg surface bacterial numbers and mortalities in the egg during incubation. Innoculation of newly-fertilized eggs with bacteria demonstrated that groups of eggs supporting high numbers of P.fluorescens suffered significantly higher mortalities during the early part of their incubation. Exposure of incubating eggs to oxolinic acid, chlortetracycline and chloramphenicol demonstrated that numbers of bacteria on egg surfaces could be significantly reduced. However, as no corresponding increase in egg hatching success was revealed, the treatment of incubating eggs with antibiotics or antimicrobial compounds can not be recommended. In commercial hatcheries bacteria are only likely to be responsible for egg deaths during incubation when environmental conditions are unfavourable. High water temperatures, slow water flow rates and high egg density all lead to increased bacterial number of egg surfaces, reduced water circulation and low levels of dissolved oxygen. Under such circumstances sufficient amounts of dissolved oxygen may not be available to support developing embryos.

Real-time biosensor for the assessment of nanotoxicity and cancer electrotherapy

Knowledge of cell electronics has led to their integration to medicine either by physically interfacing electronic devices with biological systems or by using electronics for both detection and characterization of biological materials. In this dissertation, an electrical impedance sensor (EIS) was used to measure the electrode surface impedance changes from cell samples of human and environmental toxicity of nanoscale materials in 2D and 3D cell culture models. The impedimetric response of human lung fibroblasts and rainbow trout gill epithelial cells when exposed to various nanomaterials was tested to determine their kinetic effects towards the cells and to demonstrate the biosensor's ability to monitor nanotoxicity in real-time. Further, the EIS allowed rapid, real-time and multi-sample analysis creating a versatile, noninvasive tool that is able to provide quantitative information with respect to alteration in cellular function. We then extended the application of the unique capabilities of the EIS to do real-time analysis of cancer cell response to externally applied alternating electric fields at different intermediate frequencies and low-intensity. Decreases in the growth profiles of the ovarian and breast cancer cells were observed with the application of 200 and 100 kHz, respectively, indicating specific inhibitory effects on dividing cells in culture in contrast to the non-cancerous HUVECs and mammary epithelial cells. We then sought to enhance the effects of the electric field by altering the cancer cell's electronegative membrane properties with HER2 antibody functionalized nanoparticles. An Annexin V/EthD-III assay and zeta potential were performed to determine the cell death mechanism indicating apoptosis and a decrease in zeta potential with the incorporation of the nanoparticles. With more negatively charged HER2-AuNPs attached to the cancer cell membrane, the decrease in membrane potential would thus leave the cells more vulnerable to the detrimental effects of the applied electric field due to the decrease in surface charge. Therefore, by altering the cell membrane potential, one could possibly control the fate of the cell. This whole cell-based biosensor will enhance our understanding of the responsiveness of cancer cells to electric field therapy and demonstrate potential therapeutic opportunities for electric field therapy in the treatment of cancer.

Data on the biogeochemical cycle of methane in the ocean

Geological, mineralogical and microbiological aspects of the methane cycle in water and sediments of different areas in the oceans are under consideration in the monograph. Original and published estimations of formation- and oxidation rates of methane with use of radioisotope and isotopic methods are given. The role of aerobic and anaerobic microbial oxidation of methane in production of organic matter and in formation of authigenic carbonates is considered. Particular attention is paid to processes of methane transformation in areas of its intensive input to the water column from deep-sea hydrothermal sources, mud volcanoes, and cold methane seeps.

Playwright Ntozake Shange at a pornography speak out

General note: Title and date provided by Bettye Lane.

Playwright Ntozake Shange at pornography speak out

General note: Title and date provided by Bettye Lane.

Nasalance scores for typical Irish English-speaking adults

The aim was to establish normative nasalance values for Irish English-speaking adults. Thirty men and 30 women with normal resonance read aloud 16 sentences from the Irish nasality assessment protocol, the Zoo passage, and the Rainbow passage. The speech samples were recorded using the Nasometer II 6400. Results of a mixed between–within subjects ANOVA indicated no significant gender effect on nasalance scores. The speakers showed significantly higher nasalance scores for high-pressure consonant sentences than low-pressure consonant sentences, and for the Rainbow passage than total test sentences. There was no significant difference between high-pressure consonant sentences and the Zoo passage. Compared to previous studies, the Irish young adults had lower nasalance scores than Irish children and than young adults with North American dialects.

Molecular docking and geographical information systems as tools to assess the potential impact of veterinary medicines on non-target organisms and the environment

Veterinary medicines (VMs) from agricultural industry can enter the environment in a number of ways. This includes direct exposure through aquaculture, accidental spillage and disposal, and indirect entry by leaching from manure or runoff after treatment. Many compounds used in animal treatments have ecotoxic properties that may have chronic or sometimes lethal effects when they come into contact with non-target organisms. VMs enter the environment in mixtures, potentially having additive effects. Traditional ecotoxicology tests are used to determine the lethal and sometimes reproductive effects on freshwater and terrestrial organisms. However, organisms used in ecotoxicology tests can be unrepresentative of the populations that are likely to be exposed to the compound in the environment. Most often the tests are on single compound toxicity but mixture effects may be significant and should be included in ecotoxicology testing. This work investigates the use, measured environmental concentrations (MECs) and potential impact of sea lice treatments on salmon farms in Scotland. Alternative methods for ecotoxicology testing including mixture toxicity, and the use of in silico techniques to predict the chronic impact of VMs on different species of aquatic organisms were also investigated. The Scottish Environmental Protection Agency (SEPA) provided information on the use of five sea lice treatments from 2008-2011 on Scottish salmon farms. This information was combined with the recently available data on sediment MECs for the years 2009-2012 provided by SEPA using ArcGIS 10.1. In depth analysis of this data showed that from a total of 55 sites, 30 sites had a MEC higher than the maximum allowable concentration (MAC) as set out by SEPA for emamectin benzoate and 7 sites had a higher MEC than MAC for teflubenzuron. A number of sites that were up to 16 km away from the nearest salmon farm reported as using either emamectin benzoate or teflubenzuron measured positive for the two treatments. There was no relationship between current direction and the distribution of the sea lice treatments, nor was there any evidence for alternative sources of the compounds e.g. land treatments. The sites that had MECs higher than the MAC could pose a risk to non-target organisms and disrupt the species dynamics of the area. There was evidence that some marine protected sites might be at risk of exposure to these compounds. To complement this work, effects on acute mixture toxicity of the 5 sea lice treatments, plus one major metabolite 3-phenoxybenzoic acid (3PBA), were measured using an assay using the bioluminescent bacteria Aliivibrio fischeri. When exposed to the 5 sea lice treatments and 3PBA A. fischeri showed a response to 3PBA, emamectin benzoate and azamethiphos as well as combinations of the three. In order to establish any additive effect of the sea lice treatments, the efficacy of two mixture prediction equations, concentration addition (CA) and independent action ii(IA) were tested using the results from single compound dose response curves. In this instance IA was the more effective prediction method with a linear regression confidence interval of 82.6% compared with 22.6% of CA. In silico molecular docking was carried out to predict the chronic effects of 15 VMs (including the five used as sea lice control). Molecular docking has been proposed as an alternative screening method for the chronic effects of large animal treatments on non-target organisms. Oestrogen receptor alpha (ERα) of 7 non-target bony fish and the African clawed frog Xenopus laevis were modelled using SwissModel. These models were then ‘docked’ to oestradiol, the synthetic oestrogen ethinylestradiol, two known xenoestrogens dichlorodiphenyltrichloroethane (DDT) and bisphenol A (BPA), the antioestrogen breast cancer treatment tamoxifen and 15 VMs using Auto Dock 4. Based on the results of this work, four VMs were identified as being possible xenoestrogens or anti-oestrogens; these were cypermethrin, deltamethrin, fenbendazole and teflubenzuron. Further investigation, using in vitro assays, into these four VMs has been suggested as future work. A modified recombinant yeast oestrogen screen (YES) was attempted using the cDNA of the ERα of the zebrafish Danio rerio and the rainbow trout Oncorhynchus mykiss. Due to time and difficulties in cloning protocols this work was unable to be completed. Use of such in vitro assays would allow for further investigation of the highlighted VMs into their oestrogenic potential. In conclusion, VMs used as sea lice treatments, such as teflubenzuron and emamectin benzoate may be more persistent and have a wider range in the environment than previously thought. Mixtures of sea lice treatments have been found to persist together in the environment, and effects of these mixtures on the bacteria A. fischeri can be predicted using the IA equation. Finally, molecular docking may be a suitable tool to predict chronic endocrine disrupting effects and identify varying degrees of impact on the ERα of nine species of aquatic organisms.

Standardization in the Pre-Columbian Pottery from El Caño, Panama: Specialization, Productivity and Consumption

The pottery found in the burials of El Cano is uniform in style to these made in the coclesanos valleys between 700 and 1000 AD. The coefficient of variability of the different pottery forms, evidence diverse standardizations values for polychrome and non-polychrome ceramics. Moreover, data of funerary contexts from the Cano recently excavated, suggest that elite has controlled ceramic production. This control over the production of certain goods reveals that these were important in the support or proper operational of the chiefdoms in Panama and mark the phase of splendour of this culture.

Long-term fish community response to a reach-scale stream restoration

At a global scale, aquatic ecosystems are being altered by human activities at a greater rate than at any other time in history. In recent years, grassroots efforts have generated interest in the restoration of degraded or destroyed aquatic habitats, especially small wetlands and streams where such projects are feasible with local resources. We present ecological management lessons learned from 17 years of monitoring the fish community response to the channel relocation and reach-level restoration of Juday Creek, a 3rd-order tributary of the St. Joseph River in Indiana, USA. The project was designed to increase habitat complexity, reverse the effects of accumulated fine sediment (< 2 mm diameter), and mitigate for the impacts of a new golf course development. The 1997 restoration consisted of new channel construction within two reaches of a 1.2-km section of Juday Creek that also contained two control reaches. A primary social goal of the golf course development and stream restoration was to avoid harm to the non-native brown trout fishery, as symbolic of community concerns for the watershed. Our long-term monitoring effort revealed that, although fine sediment increased over time in the restored reaches, habitat conditions have promoted the resurgence of native fish species. Since restoration, the fish assemblage has shifted from non-native Salmonidae (brown trout, rainbow trout) to native Centrarchidae (rock bass, largemouth bass, smallmouth bass). In addition, native, nongame species have remained stable or have increased in population abundance (e.g., Johnny darter, mottled sculpin). The results of this study demonstrate the value of learning from a restoration project to adjust management decisions that enhance environmental quality.

Dermoscopic variability of basal cell carcinoma according to clinical type and anatomic location

Background Correctly diagnosing basal cell carcinoma (BCC) clinical type is crucial for the therapeutic management. A systematic description of the variability of all reported BCC dermoscopic features according to clinical type and anatomic location is lacking. Objectives To describe the dermoscopic variability of BCC according to clinical type and anatomic location and to test the hypothesis of a clinical/dermoscopic continuum across superficial BCCs (sBCCs) with increasing palpability. Methods Clinical/dermoscopic images of nodular BCCs (nBCCs) and sBCCs with different degrees of palpability were retrospectively evaluated for the presence of dermoscopic criteria including degree of pigmentation, BCC-associated patterns, diverse vascular patterns, melanocytic patterns and polarized light patterns. Results We examined 501 histopathologically proven BCCs (66.9% sBCCs; 33.1% nBCCs), mainly located on trunk (46.7%; mostly sBCCs) and face (30.5%; mostly nBCCs). Short fine telangiectasias, leaf-like areas, spoke-wheel areas, small erosions and concentric structures were significantly associated with sBCC, whereas arborizing telangiectasias, blue-white veil-like structures, white shiny areas and rainbow pattern with nBCCs. Short fine telangiectasia, spoke-wheel areas and small erosions were independently associated with trunk location, whereas arborizing telangiectasias with facial location. Scalp BCCs had significantly more pigmentation and melanocytic criteria than BCCs located elsewhere. Multiple clinical/dermoscopic parameters displayed a significant linear trend across increasingly palpable sBCCs. Conclusions Particular dermoscopic criteria are independently associated with clinical type and anatomic location of BCC. Heavily pigmented, scalp BCCs are the most challenging to diagnose. A clinical/dermoscopic continuum across increasingly palpable sBCCs was detected and could be potentially important for the non-surgical management of the disease.

Biogeographical distribution of Rimicaris exoculata resident gut epibiont communities along the Mid-Atlantic Ridge hydrothermal vent sites

Rimicaris exoculata is a deep-sea hydrothermal vent shrimp which enlarged gill chamber houses a complex trophic epibiotic community. Its gut harbours an autochthonous and distinct microbial community. This species dominates hydrothermal ecosystems megafauna along the Mid-Atlantic Ridge, regardless of contrasted geochemical conditions prevailing in them. Here, the resident gut epibiont community at four contrasted hydrothermal vent sites (Rainbow/TAG/Logatchev/Ashadze) was analysed and compiled with previous data to evaluate the possible influence of site location, using 16S rRNA surveys and microscopic observations (TEM, SEM and FISH analyses). Filamentous epibionts inserted between the epithelial cells microvilli were observed on all examined samples. Results confirmed resident gut community affiliation to Deferribacteres, Mollicutes, Epsilonproteobacteria and to a lesser extent Gammaproteobacteria lineages. Still a single Deferribacteres phylotype was retrieved at all sites. Four Mollicutes-related OTUs were distinguished, one being only identified on Rainbow specimens. The topology of ribotypes median-joining networks illustrated a community diversification possibly following demographic expansions, suggesting a more ancient evolutionary history and/or a larger effective population size at Rainbow. Finally, the gill chamber community distribution was also analysed through ribotypes networks based on sequences from R. exoculata collected at Rainbow/Snake Pit/TAG/Logatchev/Ashadze sites. Results allow refining hypotheses on the epibiont role and transmission pathways.

Comparative phylogeography of the western Indian Ocean reef fauna

Assessing patterns of connectivity at the community and population levels is relevant to marine resource management and conservation. The present study reviews this issue with a focus on the western Indian Ocean (WIO) biogeographic province. This part of the Indian Ocean holds more species than expected from current models of global reef fish species richness. In this study, checklists of reef fish species were examined to determine levels of endemism in each of 10 biogeographic provinces of the Indian Ocean. Results showed that the number of endemic species was higher in the WIO than in any other region of the Indian Ocean. Endemic species from the WIO on the average had a larger body size than elsewhere in the tropical Indian Ocean. This suggests an effect of peripheral speciation, as previously documented in the Hawaiian reef fish fauna, relative to other sites in the tropical western Pacific. To explore evolutionary dynamics of species across biogeographic provinces and infer mechanisms of speciation, we present and compare the results of phylogeographic surveys based on compilations of published and unpublished mitochondrial DNA sequences for 19 Indo-Pacific reef-associated fishes (rainbow grouper Cephalopholis argus, scrawled butterflyfish Chaetodon meyeri, bluespot mullet Crenimugil sp. A, humbug damselfish Dascyllus abudafur/Dascyllus aruanus, areolate grouper Epinephelus areolatus, blacktip grouper Epinephelus fasciatus, honeycomb grouper Epinephelus merra, bluespotted cornetfish Fistularia commersonii, cleaner wrasse Labroides sp. 1, longface emperor Lethrinus sp. A, bluestripe snapper Lutjanus kasmira, unicornfishes Naso brevirosris, Naso unicornis and Naso vlamingii, blue-spotted maskray Neotrygon kuhlii, largescale mullet Planiliza macrolepis, common parrotfish Scarus psicattus, crescent grunter Terapon jarbua, whitetip reef shark Triaenodon obesus) and three coastal Indo-West Pacific invertebrates (blue seastar Linckia laevigata, spiny lobster Panulirus homarus, small giant clam Tridacna maxima). Heterogeneous and often unbalanced sampling design, paucity of data in a number of cases, and among-species discrepancy in phylogeographic structure precluded any generalization regarding phylogeographic patterns. Nevertheless, the WIO might have been a source of haplotypes in some cases and it also harboured an endemic clade in at least one case. The present survey also highlighted likely cryptic species. This may eventually affect the accuracy of the current checklists of species, which form the basis of some of the recent advances in Indo-West Pacific marine ecology and biogeography.

Patogenia de piscirickettsiosis: estudio sobre la infectividad de Piscirickettsia salmonis y el efecto de sus productos extracelulares

La piscirickettsiosis es una enfermedad contagiosa sistémica de los peces teleósteos cuyo agente causal es Piscirickettsia salmonis, una bacteria gram negativa intracelular facultativa. Esta enfermedad se ha descrito esporádicamente en distintas áreas geográficas y especies de peces en el mundo, pero es endémica y particularmente severa en salmónidos criados en agua de mar en Chile. En esta tesis se investigaron algunos aspectos de la patogénesis de esta enfermedad, estudiándose la infectividad de P. salmonis, tanto in vitro como in vivo, y buscándose además evidencias de la capacidad de secretar exotoxinas por parte de esta bacteria. Los ensayos de infectividad en células CHSE-214, procedentes de embrión de salmón chinook (Oncorhynchus tshawytscha), mostraron que existe una rápida adherencia de la bacteria a la superficie de la membrana plasmática (≤ 5 min posinoculación) seguida de su incorporación al citoplasma de estas células, proceso que ocurre entre las 3 y las 6 h posinoculación. Por su parte, el estudio de infectividad in vivo, que se realizó en trucha arcoiris (O. mykiss), reveló que este proceso comprende tres etapas principales: (i) una fase de rápida adhesión a células epiteliales principalmente de piel y branquias, pero también del canal alimentario; (ii) una invasión progresiva desde los sitios de entrada hacia tejidos más profundos hasta alcanzar el torrente sanguíneo y; (iii) una rápida diseminación vía hematógena para alcanzar virtualmente todos los tejidos corporales. Finalmente, se demostró que P. salmonis puede secretar exotoxinas termolábiles que tienen un efecto citotóxico selectivo según la célula blanco expuesta y que, probablemente, son parte de los factores de virulencia involucrados en la patogénesis de la piscirickettsiosis.

A dimensão preventiva da intervenção precoce : intervenção psicomotora na creche

O presente relatório tem como principal objetivo apresentar a caracterização do estágio realizado na Equipa Móvel de Desenvolvimento Infantil e Intervenção Precoce (EMDIIP) no âmbito da promoção do desenvolvimento infantil de crianças em contexto de creche, ao longo do segundo ano do mestrado em Reabilitação Psicomotora. Toda a intervenção psicomotora foi planeada consoante os resultados das avaliações formais, das observações informais de cada criança e das preocupações e dificuldades enunciadas pela Educadora de Infância da creche tendo sempre em conta as práticas recomendadas da Intervenção Precoce. Para além das sessões de psicomotricidade realizadas em grupo e individualmente, foi desenvolvido um trabalho de formação parental, com o intuito de promover as interações familiares e divulgar informação sobre o desenvolvimento normal das crianças, e um ajustamento do contexto da sala Arco-Íris. A psicomotricidade revelou ser uma ferramenta essencial na promoção do desenvolvimento infantil não só na sua vertente (re)educativa e terapêutica mas também na sua vertente preventiva.