Development and evaluation of the polymerase chain reaction (PCR) method for diagnosis of spring viremia of carp virus (SVCV)

Aquaculture has been expanded rapidly to become a major commercial and food-producing sector worldwide in recent decade. In parallel, viral diseases rapidly spread among farms causing enormous economic losses. The accurate detection of pathogens at early stages of infection is a key point for disease control in aquaculture. Spring Viraemia of Carp Virus (SVCV) is a very severe pathogen of carp fishes in different parts of the world and is categorized as a reportable listed disease in the annual published list of World Organization for animal Health (OIE). The objective of this study was to develop and evaluate RT- PCR test for detecting SVC virus and also the sensitivity and specificity of this test. A semi nested RT- PCR was designed using combination of three primers: two external (SVCF , SVCR) and one internal (SVCS) primers which based on conserved region of G gen. The specificity of designed primers (only external ones) by examination on Viral Hemorrhagic Septicemia Virus (VHSV) and Infectious Hematopoietic Necrosis Virus (IHNV) was confirmed. For optimizing of the PCR test, primer concentration, primer annealing temperature, cycle number and Mgcl2 concentration were surveyed. Also for validity test, prevention of false negative and Assurance of its accuracy, a competitive internal control (mimic) designed and its suitable concentration was defined. Evaluation of the sensitivity of designed test were conducted first by comparing the different commercially available RNA isolation guidelines, two guidelines: isotiocyanate phenol–chloroform based protocols (RNX–Plus Iran, Iq2000 kit Taiwan ) and two column based protocols (Cinna pure RNA Iran , high pure viral RNA kit, Roche Germany ). The results indicated that the column based protocols (Roche method and Cinna pure), yield 36.77 ng/μl and 16/47 ng/μl RNA concentration respectively, which were significantly higher than other protocols(P<0.05). Then for evaluation of extracted RNA sensitivity, Serial dilution of SVCV strain 56.70 grown in EPC (1.9×105 TCID50/ml) was examined To compare sensitivity. Extracted RNA from serial dilution with stone's primers and commercial IQ-2000 kit were examined simultaneously. The result indicated that designed semi- nested RT- PCR was able to recognize SVC virus to 10-4 dilution and stone's primer recognize to 10-3 dilution whereas Iq-2000 commercial kit did not recognized in any dilution. In high virus titer in designed test two DNA band (462 bp and 266 bp) produced, and by decreasing virus titer 462 bp was omitted. In low virus titer or lack of virus, just DNA band (mimic) 729 bp can propagate. After designing and optimizing PCR test, a total of 400 suspected cultured Cyprinus carpio with high mortality from 4 aquaculture zone of Khuzestan province were collected and tested for SVCV during 2012- 2013 using developed PCR method and IQ- 2000. The results indicated that SVC virus was not observed in samples using both methods.

Solução semi-analítica para modelagem das condições inicial e de contorno aplicáveis ao ensaio de difusão pura

Dissertação (mestrado)—Universidade de Brasília, Faculdade de Tecnologia, Departamento de Engenharia Civil e Ambiental, 2015.

Porosity imaged by a vector projection algorithm correlates with fractal dimension measured on 3D models obtained by microCT

International audience

A barreira de contacto em sujeitos toxicodependentes, através da sua expressão no Rorschach

Dissertação de Mestrado apresentada ao Instituto Superior de Psicologia Aplicada para obtenção de grau de Mestre na especialidade de Psicologia Clínica.

Climatic classification: the semi-arid tropics and its environment a review.

This paper analyses the advantages and limitations in using the Troll, Hargreaves and modified Thornthwaite approaches for the demarcation of the semi-arid tropics. Data from India, Africa, Brazil, Australia and Thailand, were used for the comparison of these three methods. The modified Thornthwaite approach provided the most relevant agriculturally oriented demarcation of the semi-arid tropics. This method in not only simple, tut uses input data that are avaliable for a global network of stations. Using this method the semi-arid tropics include major dryland or rainfed agricultural zones with annual rainfall varying from about 400 to 1,250 mm. Major dryland crops are pearl millet, sorghum, pigeonpea and groundnut. This paper also presents the brief description of climate, soils and farming systems of the semi-arid tropics.

Climate change and irrigation water requirement at Jaguaribe river basin, semi-arid northeast of Brazil.

Summary: Climate change has a potential to impact rainfall, temperature and air humidity, which have relation to plant evapotranspiration and crop water requirement. The purpose of this research is to assess climate change impacts on irrigation water demand, based on future scenarios derived from the PRECIS (Providing Regional Climates for Impacts Studies), using boundary conditions of the HadCM3 submitted to a dynamic downscaling nested to the Hadley Centre regional circulation model HadRM3P. Monthly time series for average temperature and rainfall were generated for 1961-90 (baseline) and the future (2040). The reference evapotranspiration was estimated using monthly average temperature. Projected climate change impact on irrigation water demand demonstrated to be a result of evapotranspiration and rainfall trend. Impacts were mapped over the target region by using geostatistical methods. An increase of the average crop water needs was estimated to be 18.7% and 22.2% higher for 2040 A2 and B2 scenarios, respectively. Objective ? To analyze the climate change impacts on irrigation water requirements, using downscaling techniques of a climate change model, at the river basin scale. Method: The study area was delimited between 4º39?30? and 5º40?00? South and 37º35?30? and 38º27?00? West. The crop pattern in the target area was characterized, regarding type of irrigated crops, respective areas and cropping schedules, as well as the area and type of irrigation systems adopted. The PRECIS (Providing Regional Climates for Impacts Studies) system (Jones et al., 2004) was used for generating climate predictions for the target area, using the boundary conditions of the Hadley Centre model HadCM3 (Johns et al., 2003). The considered time scale of interest for climate change impacts evaluation was the year of 2040, representing the period of 2025 to 2055. The output data from the climate model was interpolated, considering latitude/longitude, by applying ordinary kriging tools available at a Geographic Information System, in order to produce thematic maps.

Arthropoda de solo em um ecossistema semi-árido da região neotropical: composição, variabilidade temporal e estratificação

Caatinga is an important laboratory for studies about arthropods adaptations and aclimatations because its precipitation is highly variable in time. We studied the effects of time variability over the composition of Arthropods in a caatinga area. The study was carried out at a preservation area on Almas Farm, São José dos Cordeiros, Paraíba. Samples were collected in two 100 m long parallel transects, separated for a 30 m distance, in a dense tree dominated caatinga area, between August 2007 and July 2008. Samples were collected in each transect every 10 m. Ten soil samples were taken from each transect, both at 0-5 cm (A) and 5-10 cm (B) depth, resulting in 40 samples each month. The Berlese funnel method was used for fauna extraction. We registered 26 orders and the arthropods density in the soil ranged from 3237 to 22774 individuals.m-2 from January 2007 to March 2008, respectively. There was no difference between layers A and B regarding orders abundance and richness. The groups recorded include groups with few records or that had no records in the Caatinga region yet as Pauropoda, Psocoptera, Thysanoptera, Protura and Araneae. Acari was the most abundant group, with 66,7% of the total number of individuals. Soil Arthropods presented a positive correlation with soil moisture, vegetal cover, precipitation and real evapotranspiration. Increases in fauna richness and abundance were registered in February, a month after the beginning of the rainy season. A periodic rain events in arid and semiarid ecosystems triggers physiological responses in edafic organisms, like arthropods. Edafic arthropods respond to time variability in the Caatinga biome. This fauna variation has to be considered in studies of this ecosystem, because the variation of Arthropods composition in soil can affect the dynamics of the food web through time

Biologia alimentar e morfohistologia do tubo digestório do mussum, synbranchus marmoratus, bloch (osteichthyes: synbranchidae) no açude Marechal Dutra Gargalheiras , localizado no semi-árido brasileiro

Food habits and morpho-histology of the digestive tract of marbled swamp eel, Synbranchus marmoratus (Block, 1917) were investigated. The fish samples were captured during August, 2007 to July, 2008 in the Marechal Dutra reservoir, Acari, Rio Grande do Norte. The rain fall data was obtained from EMPARN. The fish captured, were measured, weighed, dissected, eviscerated and individual stomach weights were registered. The stomach contents analyses were carried out based on volumetric method, points, frequency of occurrence and applying the Index of Relative Importance. The degrees of repletion of the stomachs were determined besides the Index of Repletion relating to feeding activity variations and frequency of ingestion during the rainy and dry seasons. The rainfall varied from 0 mm a 335 mm with a mean value of 71.62 mm. Highest rainfall of 335.5 mm was registered in March, 2008 and August to December was the dry period. During the dry period the study species presented high degrees of repletion of the stomachs, with a peak value in the month of September (mean = 4.54; ± SD = 0.56). The minimum mean value of = 3.99 ± SD = 0.25 was registered in the month of May during the rainy period. The stomach contents of S. marmoratus registered show that this fish prefers animals, 78.22% of crustaceans 2.85% of mollusks, 3.25% of fish, 1.4% of insects and 13.5% of semi-digested organic matter, thus characterizing the study species as a carnivore with a preference for crustaceans. The morpho-histological aspects of the digestive tract of S. marmoratus indicate that the mouth is terminal adapted to open widely, thin lips with taste buds, small villiform teeth forming a single series on maxillas, four pairs of branchial arches with short and widely spaced branchial rays. The oesophagus is short and cylindrical with a small diameter. The oesophagus wall is thick with mucas surface and internal parallel folds. The stomach is retilinical in form, presenting cardiac, caecal and pyloric portions. The caecal portion is long and is intermediary in position between the cardiac and pyloric portions. The cardiac portion of the stomach is short and cylindrical formed of simple epithelial cylindrical mucus cells. The caecal portion is long with narrow walls, a big cavity and smaller folds which give rise to gastric glands. The phyloric portion has no glands and primary or secondary mucas folds. The morphohistological aspects of the digestive tract of S. marmoratus indicate its adaptation to a carnivorous feeding habit

Semi-pelagic longline selectivity for two demersal species from the Azores: the black spot sea bream (Pagellus bogaraveo) and the bluemouth rockfish (Helicolenus dactylopterus dactylopterus)

The selectivity of four hook sizes (STELL brand(1), Quality 2335, numbers 12, 9, 6 and 4) used in a semi-pelagic longline fishery was studied in the Azores. Two species were caught in sufficient numbers for modelling of selectivity: the black spot sea bream (Pagellus bogaraveo) and the bluemouth rockfish (Helicolenus dactylopterus dactylopterus). A maximum likelihood method was used to fit a versatile model which can be used to describe a wide range of selectivity curves; from bell-shaped to asymptotic. Significant differences in size selectivity between hooks were found for both species. In the case of Pagellus bogaraveo, the smallest hook (number 12) had the lowest catch rates and all hooks were characterised by logistic-type selectivity curves, with sizes at 50% selectivity of: 27.9, 30.4, and 32.8 cm for hooks numbers 12, 9 and 6, respectively. The number 9 hook was the most efficient for Helicolenus d. dactylopterus, with selectivity curves varying from strongly skewed to the right for the number 12 hook to logistic-type for the numbers 6 and 4 hooks. Sizes at 50% selectivity for this species were 16.8, 18.7, 20.7, and 22.0 cm. respectively. (C) 1999 Elsevier Science B.V. All rights reserved.

Query routing in cooperative semi-structured peer-to-peer information retrieval networks

Conventional web search engines are centralised in that a single entity crawls and indexes the documents selected for future retrieval, and the relevance models used to determine which documents are relevant to a given user query. As a result, these search engines suffer from several technical drawbacks such as handling scale, timeliness and reliability, in addition to ethical concerns such as commercial manipulation and information censorship. Alleviating the need to rely entirely on a single entity, Peer-to-Peer (P2P) Information Retrieval (IR) has been proposed as a solution, as it distributes the functional components of a web search engine – from crawling and indexing documents, to query processing – across the network of users (or, peers) who use the search engine. This strategy for constructing an IR system poses several efficiency and effectiveness challenges which have been identified in past work. Accordingly, this thesis makes several contributions towards advancing the state of the art in P2P-IR effectiveness by improving the query processing and relevance scoring aspects of a P2P web search. Federated search systems are a form of distributed information retrieval model that route the user’s information need, formulated as a query, to distributed resources and merge the retrieved result lists into a final list. P2P-IR networks are one form of federated search in routing queries and merging result among participating peers. The query is propagated through disseminated nodes to hit the peers that are most likely to contain relevant documents, then the retrieved result lists are merged at different points along the path from the relevant peers to the query initializer (or namely, customer). However, query routing in P2P-IR networks is considered as one of the major challenges and critical part in P2P-IR networks; as the relevant peers might be lost in low-quality peer selection while executing the query routing, and inevitably lead to less effective retrieval results. This motivates this thesis to study and propose query routing techniques to improve retrieval quality in such networks. Cluster-based semi-structured P2P-IR networks exploit the cluster hypothesis to organise the peers into similar semantic clusters where each such semantic cluster is managed by super-peers. In this thesis, I construct three semi-structured P2P-IR models and examine their retrieval effectiveness. I also leverage the cluster centroids at the super-peer level as content representations gathered from cooperative peers to propose a query routing approach called Inverted PeerCluster Index (IPI) that simulates the conventional inverted index of the centralised corpus to organise the statistics of peers’ terms. The results show a competitive retrieval quality in comparison to baseline approaches. Furthermore, I study the applicability of using the conventional Information Retrieval models as peer selection approaches where each peer can be considered as a big document of documents. The experimental evaluation shows comparative and significant results and explains that document retrieval methods are very effective for peer selection that brings back the analogy between documents and peers. Additionally, Learning to Rank (LtR) algorithms are exploited to build a learned classifier for peer ranking at the super-peer level. The experiments show significant results with state-of-the-art resource selection methods and competitive results to corresponding classification-based approaches. Finally, I propose reputation-based query routing approaches that exploit the idea of providing feedback on a specific item in the social community networks and manage it for future decision-making. The system monitors users’ behaviours when they click or download documents from the final ranked list as implicit feedback and mines the given information to build a reputation-based data structure. The data structure is used to score peers and then rank them for query routing. I conduct a set of experiments to cover various scenarios including noisy feedback information (i.e, providing positive feedback on non-relevant documents) to examine the robustness of reputation-based approaches. The empirical evaluation shows significant results in almost all measurement metrics with approximate improvement more than 56% compared to baseline approaches. Thus, based on the results, if one were to choose one technique, reputation-based approaches are clearly the natural choices which also can be deployed on any P2P network.

Congenital thoracic vertebral malformations in brachycephalic “screw-tailed” dog breeds: Validation of a human classification scheme and a method of Cobb angle measurement in the assessment of vertebral column deformity

Congenital vertebral malformations are common in brachycephalic “screw-tailed” dog breeds such as French bulldogs, English bulldogs, Boston terriers, and Pugs. Those vertebral malformations disrupt the normal vertebral column anatomy and biomechanics, potentially leading to deformity of the vertebral column and subsequent neurological dysfunction. The initial aim of this work was to study and determine whether the congenital vertebral malformations identified in those breeds could be translated in a radiographic classification scheme used in humans to give an improved classification, with clear and well-defined terminology, with the expectation that this would facilitate future study and clinical management in the veterinary field. Therefore, two observers who were blinded to the neurologic status of the dogs classified each vertebral malformation based on the human classification scheme of McMaster and were able to translate them successfully into a new classification scheme for veterinary use. The following aim was to assess the nature and the impact of vertebral column deformity engendered by those congenital vertebral malformations in the target breeds. As no gold standard exists in veterinary medicine for the calculation of the degree of deformity, it was elected to adapt the human equivalent, termed the Cobb angle, as a potential standard reference tool for use in veterinary practice. For the validation of the Cobb angle measurement method, a computerised semi-automatic technique was used and assessed by multiple independent observers. They observed not only that Kyphosis was the most common vertebral column deformity but also that patients with such deformity were found to be more likely to suffer from neurological deficits, more especially if their Cobb angle was above 35 degrees.

Quantification of free auxins in semi-hardwood plant cuttings and microshoots by dispersive liquid–liquid microextraction/microwave derivatization and GC/MS analysis

Several studies have suggested that differences in the natural rooting ability of plant cuttings could be attributed to differences in endogenous auxin levels. Hence, during rooting experiments, it is important to be able to routinely monitor the evolution of endogenous levels of plant hormones. This work reports the development of a new method for the quantification of free auxins in auxin-treated Olea europaea (L.) explants, using dispersive liquid–liquid microextraction (DLLME) and microwave assisted derivatization (MAD) followed by gas chromatography/mass spectrometry (GC/MS) analysis. Linear ranges of 0.5–500 ng mL 1 and 1–500 mg mL 1 were used for the quantification of indole-3-acetic acid (IAA) and indole-3-butyric acid (IBA), respectively. Determined by serial dilutions, the limits of detection (LOD) and quantification (LOQ) were 0.05 ng mL 1 and 0.25 ng mL 1, respectively for both compounds. When using the calibration curve for determination, the LOQ corresponded to 0.5 ng mL 1 (IAA) and 0.5 mg mL 1 (IBA). The proposed method proved to be substantially faster than other alternatives, and allowed free auxin quantification in real samples of semi-hardwood cuttings and microshoots of two olive cultivars. The concentrations found in the analyzed samples are in the range of 0.131–0.342 mg g 1 (IAA) and 20–264 mg g 1 (IBA).

Comparison of Explicit and Implicit Methods of Cross-Cultural Learning in an International Classroom

The paper addresses a gap in the literature concerning the difference between enhanced and not enhanced cross-cultural learning in an international classroom. The objective of the described research was to clarify if the environment of international classrooms could enhance cross-cultural competences significantly enough or if additional focus on cross-cultural learning as an explicit objective of learning activities would add substantially to the experience. The research question was defined as “how can a specific exercise focused on cross-cultural learning enhance the cross-cultural skills of university students in an international classroom?”. Surveys were conducted among interna- tional students in three leading Central-European Universities in Lithuania, Poland and Hungary to measure the increase of their cross-cultural competences. The Lithuanian and Polish classes were composed of international students and concentrated on International Management/Business topics (explicit method). The Hungarian survey was done in a general business class that just happened to be international in its composition (implicit method). Overall, our findings prove that the implicit method resulted in comparable, somewhat even stronger effectiveness than the explicit method. The study method included the analyses of students’ individual increases in each study dimension and construction of a compound measure to note the overall results. Our findings confirm the power of the international classroom as a stimulating environment for latent cross-cultural learning even without specific exercises focused on cross-cultural learning itself. However, the specific exercise did induce additional learning, especially related to cross-cultural awareness and communication with representatives of other cultures, even though the extent of that learning may be interpreted as underwhelming. The main conclusion from the study is that the diversity of the students engaged in a project provided an environment that supported cross-cultural learning, even without specific culture-focused reflections or exercises.

A Highly Improved Method For Sensitive Determination Of Amitriptyline In Pharmaceutical Formulations Using An Unmodified Carbon Nanotube Electrode In The Presence Of Sulfuric Acid.

The present paper describes a novel, simple and reliable differential pulse voltammetric method for determining amitriptyline (AMT) in pharmaceutical formulations. It has been described for many authors that this antidepressant is electrochemically inactive at carbon electrodes. However, the procedure proposed herein consisted in electrochemically oxidizing AMT at an unmodified carbon nanotube paste electrode in the presence of 0.1 mol L(-1) sulfuric acid used as electrolyte. At such concentration, the acid facilitated the AMT electroxidation through one-electron transfer at 1.33 V vs. Ag/AgCl, as observed by the augmentation of peak current. Concerning optimized conditions (modulation time 5 ms, scan rate 90 mV s(-1), and pulse amplitude 120 mV) a linear calibration curve was constructed in the range of 0.0-30.0 μmol L(-1), with a correlation coefficient of 0.9991 and a limit of detection of 1.61 μmol L(-1). The procedure was successfully validated for intra- and inter-day precision and accuracy. Moreover, its feasibility was assessed through analysis of commercial pharmaceutical formulations and it has been compared to the UV-vis spectrophotometric method used as standard analytical technique recommended by the Brazilian Pharmacopoeia.