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.

Automated, objective texture segmentation of multibeam echosounder data - Seafloor survey and substrate maps from James Island to Ozette Lake, Washington Outer Coast

Without knowledge of basic seafloor characteristics, the ability to address any number of critical marine and/or coastal management issues is diminished. For example, management and conservation of essential fish habitat (EFH), a requirement mandated by federally guided fishery management plans (FMPs), requires among other things a description of habitats for federally managed species. Although the list of attributes important to habitat are numerous, the ability to efficiently and effectively describe many, and especially at the scales required, does not exist with the tools currently available. However, several characteristics of seafloor morphology are readily obtainable at multiple scales and can serve as useful descriptors of habitat. Recent advancements in acoustic technology, such as multibeam echosounding (MBES), can provide remote indication of surficial sediment properties such as texture, hardness, or roughness, and further permit highly detailed renderings of seafloor morphology. With acoustic-based surveys providing a relatively efficient method for data acquisition, there exists a need for efficient and reproducible automated segmentation routines to process the data. Using MBES data collected by the Olympic Coast National Marine Sanctuary (OCNMS), and through a contracted seafloor survey, we expanded on the techniques of Cutter et al. (2003) to describe an objective repeatable process that uses parameterized local Fourier histogram (LFH) texture features to automate segmentation of surficial sediments from acoustic imagery using a maximum likelihood decision rule. Sonar signatures and classification performance were evaluated using video imagery obtained from a towed camera sled. Segmented raster images were converted to polygon features and attributed using a hierarchical deep-water marine benthic classification scheme (Greene et al. 1999) for use in a geographical information system (GIS). (PDF contains 41 pages.)

Diagnosis and the management constituency of small-scale fisheries

Diagnosis and adaptive management can help improve the ability of small-scale fisheries (SSF) in the developing world to better cope with and adapt to both external drivers and internal sources of uncertainty. This paper presents a framework for diagnosis and adaptive management and discusses ways of implementing the first two phases of learning: diagnosis and mobilising an appropriate management constituency. The discussion addresses key issues and suggests suitable approaches and tools as well as numerous sources of further information. Diagnosis of a SSF defines the system to be managed, outlines the scope of the management problem in terms of threats and opportunities, and aims to construct realistic and desired future projections for the fishery. These steps can clarify objectives and lead to development of indicators necessary for adaptive management. Before management, however, it is important to mobilize a management constituency to enact change. Ways of identifying stakeholders and understanding both enabling and obstructive interactions and management structures are outlined. These preliminary learning phases for adaptive SSF management are expected to work best if legitimised by collaborative discussion among fishery stakeholders drawing on multiple knowledge systems and participatory approaches to assessment. (PDF contains 33 pages)

A review of diseases associated with cage culture systems: diagnosis and control in smale scale fish farming

Fish cage culture is a rapid aquacultural practice of producing fish with more yield compared to traditional pond culture. Several species cultured by this method include Cyprinus carpio, Orechromis niloticus, Sarotherodon galilaeus, Tilapia zilli, Clarias lazera, C. gariepinus, Heterobranchus bidorsalis, Citharinus citharus, Distochodus rostratus and Alestes dentes. However, the culture of fish in cages has some problems that are due to mechanical defects of the cage or diseases due to infection. The mechanical problems which may lead to clogged net, toxicity and easy access by predators depend on defects associated with various types of nets which include fold sieve cloth net, wire net, polypropylene net, nylon, galvanized and welded net. The diseases problems are of two types namely introduced diseases due to parasites. The introduced parasites include Crustaseans, Ergasilus sp. Argulus africana, and Lamprolegna sp, Helminth, Diplostomulum tregnna: Protozoan, Trichodina sp, Myxosoma sp, Myxobolus sp. the second disease problems are inherent diseases aggravated by the very rich nutrient environment in cages for rapid bacterial, saprophytic fungi, and phytoplanktonic bloom resulting in clogging of net, stagnation of water and low biological oxygen demand (BOD). The consequence is fish kill, prevalence of gill rot and dropsy conditions. Recommendations on routine cage hygiene, diagnosis and control procedures to reduce fish mortality are highlighted

Molecular probes and polymerase chain reaction (PCR) -based kits for diagnosis of shrimp diseases

Technology for effective and fast diagnosis of animal diseases is essential for developing aquaculture management strategies. This paper reviews the conventional techniques for shrimp disease diagnosis and discusses the emergence of nuclei acid probes and polymerase chain reaction (PCR)-based kits as powerful tools for rapid and accurate detection of shrimp diseases.

A simple and objective bioeconomic index for choosing species for culture

Details are given of the development of a bioeconomic culture index that may be used to assist in choosing fish for culture. This index takes into account growth performance indexes and mean annual prices of the fish. Examples are given of the culture indices of some of Kuwait's commercially important fish species. The proposed indices given realistic output for these fish and allow comparison and evaluation of different taxa with widely differing biological and economic characteristics.

An objective classification of climatic regions in the Pacific and Indian oceans

We have applied a number of objective statistical techniques to define homogeneous climatic regions for the Pacific Ocean, using COADS (Woodruff et al 1987) monthly sea surface temperature (SST) for 1950-1989 as the key variable. The basic data comprised all global 4°x4° latitude/longitude boxes with enough data available to yield reliable long-term means of monthly mean SST. An R-mode principal components analysis of these data, following a technique first used by Stidd (1967), yields information about harmonics of the annual cycles of SST. We used the spatial coefficients (one for each 4-degree box and eigenvector) as input to a K-means cluster analysis to classify the gridbox SST data into 34 global regions, in which 20 comprise the Pacific and Indian oceans. Seasonal time series were then produced for each of these regions. For comparison purposes, the variance spectrum of each regional anomaly time series was calculated. Most of the significant spectral peaks occur near the biennial (2.1-2.2 years) and ENSO (~3-6 years) time scales in the tropical regions. Decadal scale fluctuations are important in the mid-latitude ocean regions.

A sudden collapse in distribution of Pacific sardine (Sardinops sagax) off southwestern Australia enables an objective re-assessment of biomass estimates

Fishery-independent estimates of spawning biomass (BSP) of the Pacific sardine (Sardinops sagax) on the south and lower west coasts of Western Australia (WA) were obtained periodically between 1991 and 1999 by using the daily egg production method (DEPM). Ichthyoplankton data collected during these surveys, specifically the presence or absence of S. sagax eggs, were used to investigate trends in the spawning area of S. sagax within each of four regions. The expectation was that trends in BSP and spawning area were positively related. With the DEPM model, estimates of BSP will change proportionally with spawning area if all other variables remain constant. The proportion of positive stations (PPS), i.e., stations with nonzero egg counts — an objective estimator of spawning area — was high for all south coast regions during the early 1990s (a period when the estimated BSP was also high) and then decreased after the mid-1990s. There was a decrease in PPS from the mid-1990s to 1999. The particularly low estimates in 1999 followed a severe epidemic mass mortality of S. sagax throughout their range across southern Australia. Deviations from the expected relationship between BSP and PPS were used to identify uncertainty around estimates of BSP. Because estimation of spawning area is subject to less sampling bias than estimation of BSP, the deviation in the relation between the two provides an objective basis for adjusting some estimates of the latter. Such an approach is particularly useful for fisheries management purposes when sampling problems are suspected to be present. The analysis of PPS undertaken from the same set of samples from which the DEPM estimate is derived will help provide information for stock assessments and for the management of purse-seine fisheries.

Simulation and diagnosis of large-scale atmospheric moisture and surface hydrology over North America

We describe a preliminary investigation into large-scale atmospheric and surface moisture variations over North America. We compare large-scale hydrologic budgets in the Los Alamos general circulation model (GCM) to observed precipitation and vertically integrated atmospheric moisture fluxes derived from the National Meteorological Center's operational analyses. THe GCM faithfully simulates the integrated flux divergence and P-E differences. However, the integrated moisture content is too low, and precipitation and evaporation are too high. The model produces summertime soil moisture dryness, which supports previous studies showing increased droughts under warmer conditions.

An objective method to monitor and quantify texture of salted herring during ripening in barrels at 4°C

Changes in the texture (elastic nature) of the flesh of barrel salted herring during the ripening process at 4°C have been monitored. The method employs the analysis of stress-relaxation curves after compression to half of the sample thickness on an lnstron Model 1112. The parameter 'T/P' for each sample represents the reciprocal of the gradient of a line connecting P and T0.368p. This parameter characteristic of each sample's texture was calculated as the ratio of 'T/P' where, T is the relaxation time and is defined as the time required for a stress at constant strain to decrease to 1/e of its original value, where 'e' is the base of natural logarithms (2.7183). Since 1/e=0.368, the relaxation time is the time required for the force to decay to 36.8% of its original value. P is the peak height of the curve (i.e. the force value at the maximum height). This method was adopted from the bakery industry for testing the degree of gluten development in bread dough. The 'T/P' values obtained over the course of ripening for differently treated salted-herring in barrels ranged between 1 and 12. The trends in 'T/P' value, during ripening period for the different samples, appeared to be parallel changes in texture perceived by sensory observation (subjective measurement), although the heterogeneous nature of the samples gave standard deviations, about the replicate sample mean, around 5%. The method appears promising as an objective measure for monitoring this aspect of the textural quality of barrel salted-herring through ripening if reproducibility of test results can be improved by more careful standardization of sample preparation and test protocol.