Emergence of carp edema virus (CEV) and its significance to European common carp and koi Cyprinus carpio.

Carp edema virus disease (CEVD), also known as koi sleepy disease, is caused by a poxvirus associated with outbreaks of clinical disease in koi and common carp Cyprinus carpio. Originally characterised in Japan in the 1970s, international trade in koi has led to the spread of CEV, although the first recognised outbreak of the disease outside of Japan was not reported until 1996 in the USA. In Europe, the disease was first recognised in 2009 and, as detection and diagnosis have improved, more EU member states have reported CEV associated with disease outbreaks. Although the structure of the CEV genome is not yet elucidated, molecular epidemiology studies have suggested distinct geographical populations of CEV infecting both koi and common carp. Detection and identification of cases of CEVD in common carp were unreliable using the original PCR primers. New primers for conventional and quantitative PCR (qPCR) have been designed that improve detection, and their sequences are provided in this paper. The qPCR primers have successfully detected CEV DNA in archive material from investigations of unexplained carp mortalities conducted >15 yr ago. Improvement in disease management and control is possible, and the principles of biosecurity, good health management and disease surveillance, applied to koi herpesvirus disease, can be equally applied to CEVD. However, further research studies are needed to fill the knowledge gaps in the disease pathogenesis and epidemiology that, currently, prevent an accurate assessment of the likely impact of CEVD on European koi and common carp aquaculture and on wild carp stocks.

Validation of a serum neutralization test for detection of antibodies specific to cyprinid herpesvirus 3 in infected common and koi carp (Cyprinus carpio).

Cyprinid herpesvirus 3 (CyHV-3) is the aetiological agent of a serious infective, notifiable disease affecting common carp and varieties. In survivors, infection is generally characterized by a subclinical latency phase with restricted viral replication. The CyHV-3 genome is difficult to detect in such carrier fish that represent a potential source of dissemination if viral reactivation occurs. In this study, the analytical and diagnostic performance of an alternative serum neutralization (SN) method based on the detection of CyHV-3-specific antibodies was assessed using 151 serum or plasma samples from healthy and naturally or experimentally CyHV-3-infected carp. French CyHV-3 isolate 07/108b was neutralized efficiently by sera from carp infected with European, American and Taiwanese CyHV-3 isolates, but no neutralization was observed using sera specific to other aquatic herpesviruses. Diagnostic sensitivity, diagnostic specificity and repeatability of 95.9%, 99.0% and 99.3%, respectively, were obtained, as well as a compliance rate of 89.9% in reproducibility testing. Neutralizing antibodies were steadily detected in infected carp subjected to restrictive or permissive temperature variations over more than 25 months post-infection. The results suggest that this non-lethal diagnostic test could be used in the future to improve the epidemiological surveillance and control of CyHV-3 disease.

Carp edema virus in Polish aquaculture - evidence of significant sequence divergence and a new lineage in common carp Cyprinus carpio (L.).

Fish samples initially collected by local veterinarians on the common and koi carp farms in Poland between 2013 and 2015 as part of a KHV surveillance programme, when the water temperature was between 16 and 26 °C, and were also tested for CEV by qPCR. A partial 478 nucleotide fragment of the 4a gene was subsequently generated from 17 qPCR-positive common carp Cyprinus carpio samples from 36 farm sites tested during the period. Sequence alignments and analysis revealed the presence of CEV in Poland both in common carp as well as in koi carp farms, and phylogenetic analysis assigned the Polish CEV sequences into three distinct genogroups. A lineage which includes the original sequences obtained from koi carp in Japan (genogroup II) included sequences from both koi carp and common carp, and the second lineage (genogroup I) contained sequences from common carp only. A third lineage (genogroup III) which was more closely related to the genogroup II also consisted of sequences from common carp only. The latter represents a lineage of CEV not previously described in the literature.

Validation of a KHV antibody enzyme-linked immunosorbent assay (ELISA).

Koi herpesvirus (KHV) causes KHV disease (KHVD). The virus is highly contagious in carp or koi and can induce a high mortality. Latency and, in some cases, a lack of signs presents a challenge for virus detection. Appropriate immunological detection methods for anti-KHV antibodies have not yet been fully validated for KHV. Therefore, it was developed and validated an enzyme-linked immunosorbent assay (ELISA) to detect KHV antibodies. The assay was optimized with respect to plates, buffers, antigens and assay conditions. It demonstrated high diagnostic and analytical sensitivity and specificity and was particularly useful at the pond or farm levels. Considering the scale of the carp and koi industry worldwide, this assay represents an important practical tool for the indirect detection of KHV, also in the absence of clinical signs.

Isotropic round-wire multifilament cuprate superconductor for generation of magnetic fields above 30 T.

Magnets are the principal market for superconductors, but making attractive conductors out of the high-temperature cuprate superconductors (HTSs) has proved difficult because of the presence of high-angle grain boundaries that are generally believed to lower the critical current density, J(c). To minimize such grain boundary obstacles, HTS conductors such as REBa2Cu3O(7-x) and (Bi, Pb)2Sr2Ca2Cu3O(10-x) are both made as tapes with a high aspect ratio and a large superconducting anisotropy. Here we report that Bi2Sr2CaCu2O(8-x) (Bi-2212) can be made in the much more desirable isotropic, round-wire, multifilament form that can be wound or cabled into arbitrary geometries and will be especially valuable for high-field NMR magnets beyond the present 1 GHz proton resonance limit of Nb3Sn technology. An appealing attribute of this Bi-2212 conductor is that, being without macroscopic texture, it contains many high-angle grain boundaries but nevertheless attains a very high J(c) of 2,500 A mm(-2) at 20 T and 4.2 K. The large potential of the conductor has been demonstrated by building a small coil that generated almost 2.6 T in a 31 T background field. This demonstration that grain boundary limits to high Jc can be practically overcome underlines the value of a renewed focus on grain boundary properties in non-ideal geometries.

First isolation of hirame rhabdovirus from freshwater fish in Europe.

A rhabdovirus was isolated in cell culture inoculated with tissue material from diseased grayling, Thymallus thymallus (L.), originating from a fish farm affected by a mortality episode in Poland. Diagnostics tests showed that the virus was not related to novirhabdoviruses known in Europe, nor to vesiculovirus-like species, except perch rhabdovirus (PRhV) with which it shared moderate serological relations. However, RT-PCR with PRhV probes gave negative results. To identify the virus, a random-priming sequence-independent single primer amplification was adopted. Surprisingly, two of the obtained sequences exhibited a high identity (>99%) with hirame rhabdovirus (HIRRV), a novirhabdovirus usually found in fish in marine Asiatic countries, for instance Japan, China and Korea. The full-length sequence of the phosphoprotein gene (P) demonstrated a higher identity of the present isolate with HIRRV from China compared with the Korean isolate. An identical viral sequence was also found in brown trout, Salmo trutta trutta L., affected by mortalities in a second farm in the same region, after a likely contamination from the grayling farm. To our knowledge, this is the first report of HIRRV in Europe, and in two hosts from fresh water that have not been described before as susceptible species.

Trade practices are main factors involved in the transmission of viral haemorrhagic septicaemia.

Viral haemorrhagic septicaemia (VHS), caused by the novirhabdovirus viral haemorrhagic septicaemia virus (VHSV), causes significant economic problems to European rainbow trout, Oncorhynchus mykiss (Walbaum), production. The virus isolates can be divided into four distinct genotypes with additional subgroups. The main source of outbreaks in European rainbow trout farming is sublineage Ia isolates. Recently, this group of isolates has been further subdivided in to two subclades of which the Ia-2 consists of isolates occurring mainly in Continental Europe outside of Denmark. In this study, we sequenced the full-length G-gene sequences of 24 VHSV isolates that caused VHS outbreaks in Polish trout farms between 2005 and 2009. All these isolates were identified as genotype Ia-2; they divided however into two genetically distinct subgroups, that we name Pol I and Pol II. The Pol I isolates mainly caused outbreaks in the southern part of Poland, while Pol II isolates predominantly were sampled in the north of Poland, although it seems that they have been transmitted to other parts of the country. Molecular epidemiology was used for characterization of transmission pathways. This study shows that a main cause of virus transmission appears to be movement of fish. At least in Polish circumstances trading practices appear to have significant impact on spreading of VHSV infection.

Process to densify Bi2Sr2CaCu2Ox round wire with overpressure before coil winding and final overpressure heat treatment.

Overpressure (OP) processing of wind-and-react Bi2Sr2CaCu2Ox (2212) round wire compresses the wire to almost full density, decreasing its diameter by about 4 % without change in wire length and substantially raising its J c . However, such shrinkage can degrade coil winding pack density and magnetic field homogeneity. To address this issue, we here present an overpressure predensification (OP-PD) heat treatment process performed before melting the 2212, which greatly reduces wire diameter shrinkage during the full OP heat treatment (OP-HT). We found that about 80 % of the total wire diameter shrinkage occurs during the 50 atm OP-PD before melting. We successfully wound such pre-densified 1.2 mm diameter wires onto coil mandrels as small as 10 mm diameter for Ag-Mg-sheathed wire and 5 mm for Ag-sheathed wire, even though such small diameters impose plastic strains up to 12% on the conductor. A further ~20% shrinkage occurred during a standard OP-HT. No 2212 leakage was observed for coil diameters as small as 20 mm for Ag-Mg-sheathed wire and 10 mm for Ag-sheathed wire, and no J c degradation was observed on straight samples and 30 mm diameter coils.

Effect of sheath material and reaction overpressure on Ag extrusions into the TiO2 insulation coating of Bi-2212 round wire.

In order to develop a high current density in coils, Bi-2212 wires must be electrically discrete in tight winding packs. It is vital to use an insulating layer that is thin, fulfils the dielectric requirements, and can survive the heat treatment whose maximum temperature reaches 890 °C. A thin (20-30 µm) ceramic coating could be better as the insulating layer compared to alumino-silicate braided fiber insulation, which is about 100 µm thick and reacts with the Ag sheath during heat treatment, degrading the critical current density (Jc). At present, TiO2 seems to be the most viable ceramic material for such a thin insulation because it is chemically compatible with Ag and Bi-2212 and its sintering temperature is lower than the maximum temperature used for the Bi-2212 heat treatment. However, recent tests of a large Bi-2212 coil insulated only with TiO2 showed severe electrical shorting between the wires after over pressure heat treatment (OPHT). The origin of the shorting was frequent silver extrusions that penetrated the porous TiO2 layer and electrically connected adjacent Bi-2212 wires. To understand the mechanism of this unexpected behaviour, we investigated the effect of sheath material and hydrostatic pressure on the formation of Ag extrusions. We found that Ag extrusions occur only when TiO2-insulated Ag-0.2%Mg sheathed wire (Ag(Mg) wire) undergoes OPHT at 50 bar. No Ag extrusions were observed when the TiO2-insulated Ag(Mg) wire was processed at 1 bar. The TiO2-insulated wires sheathed with pure Ag that underwent 50 bar OPHT were also free from Ag extrusions. A key finding is that the Ag extrusions emanating from the Ag(Mg) sheath actually contain no MgO, suggesting that local depletion of MgO facilitates local, heterogeneous deformation of the sheath under hydrostatic overpressure. Our study also suggests that predensifying the Ag(Mg) wire before insulating it with TiO2 and doing the final OPHT can potentially prevent Ag extrusion.

Understanding the densification process of Bi2Sr2CaCu2Ox round wires with overpressure processing and its effect on critical current density.

Overpressure (OP) processing increases the critical current density (JC ) of Bi2Sr2CaCu2Ox (2212) round wires by shrinking the surrounding Ag matrix around the 2212 filaments, driving them close to full density and greatly increasing the 2212 grain connectivity. Indeed densification is vital for attaining the highest JC . Here, we investigate the time and temperature dependence of the wire densification. We find that the wire diameter decreases by 3.8 ± 0.3 % after full heat treatment at 50 atm and 100 atm OP. At 50 atm OP pressure, the filaments start densifying above 700 °C and reach a 3.30 ± 0.07 % smaller diameter after 2 h at 820 °C, which is below the melting point of 2212 powder. The densification is homogeneous and does not change the filament shape before melting. The growth of non-superconducting phases is observed at 820 °C, suggesting that time should be minimized at high temperature prior to melting the 2212 powder. Study of an open-ended 2.2 m long wire sample shows that full densification and the high OP JC (JC varies by about 3.1 times over the 2.2 m long wire) is reached about 1 m from the open ends, thus showing that coil-length wires can be protected from leaky seals by adding at least 1 m of sacrificial wire at each end.

Comparison of growth texture in round Bi2212 and flat Bi2223 wires and its relation to high critical current density development.

Why Bi(2)Sr(2)CaCu(2)Ox (Bi2212) allows high critical current density Jc in round wires rather than only in the anisotropic tape form demanded by all other high temperature superconductors is important for future magnet applications. Here we compare the local texture of state-of-the-art Bi2212 and Bi2223 ((Bi,Pb)(2)Sr(2)Ca(2)Cu(3)O(10)), finding that round wire Bi2212 generates a dominant a-axis growth texture that also enforces a local biaxial texture (FWHM <15°) while simultaneously allowing the c-axes of its polycrystals to rotate azimuthally along and about the filament axis so as to generate macroscopically isotropic behavior. By contrast Bi2223 shows only a uniaxial (FWHM <15°) c-axis texture perpendicular to the tape plane without any in-plane texture. Consistent with these observations, a marked, field-increasing, field-decreasing J(c)(H) hysteresis characteristic of weak-linked systems appears in Bi2223 but is absent in Bi2212 round wire. Growth-induced texture on cooling from the melt step of the Bi2212 J(c) optimization process appears to be the key step in generating this highly desirable microstructure.