Traits of a mussel transmissible cancer are reminiscent of a parasitic life style.

Some cancers have evolved the ability to spread from host to host by transmission of cancerous cells. These rare biological entities can be considered parasites with a host-related genome. Still, we know little about their specific adaptation to a parasitic lifestyle. MtrBTN2 is one of the few lineages of transmissible cancers known in the animal kingdom. Reported worldwide, MtrBTN2 infects marine mussels. We isolated MtrBTN2 cells circulating in the hemolymph of cancerous mussels and investigated their phenotypic traits. We found that MtrBTN2 cells had remarkable survival capacities in seawater, much higher than normal hemocytes. With almost 100% cell survival over three days, they increase significantly their chances to infect neighboring hosts. MtrBTN2 also triggered an aggressive cancerous process: proliferation in mussels was ~ 17 times higher than normal hemocytes (mean doubling time of ~ 3 days), thereby favoring a rapid increase of intra-host population size. MtrBTN2 appears to induce host castration, thereby favoring resources re-allocation to the parasites and increasing the host carrying capacity. Altogether, our results highlight a series of traits of MtrBTN2 consistent with a marine parasitic lifestyle that may have contributed to the success of its persistence and dissemination in different mussel populations across the globe.

Implementation of various approaches to study the prevalence, incidence and progression of disseminated neoplasia in mussel stocks.

Marine mussel production is of substantial economic interest in numerous coastal areas worldwide, making crucial the study of pathologies that affect them. Disseminated neoplasia (DN) has recently been suggested to be linked to blue mussel, Mytilus edulis, mortality outbreaks observed in France since 2014, although the evidence remains indirect. In order to improve DN detection and monitoring, we compared the sensitivity of four diagnostic tools, namely haemocytology, histology, flow cytometry, and genetics. Haemocytological examination gave the best results in sensitivity and had the advantage of being non-invasive, allowing disease progression to be followed in affected mussels. Using this approach, we showed that DN progression is usually slow, and we provide evidence of remission events. We observed a high diversity of forms and mitotic features of neoplastic cells located in the vesicular connective tissue but rarely in the haemolymph. Circulating cells occur as four main types but are homogenous in morphology and DNA content within a single individual. Polyploidy proved very high, from 8 N to 18 N. Genetic analysis of haemolymph DNA showed that a Mytilus trossulus genetic signal was associated with almost all the DN cases here diagnosed by haemocytological examination, regardless of the DN type. This result corroborates DN is a transmissible cancer that first originated in a M. trossulus host and subsequently crossed into M. edulis. No pre-neoplastic conditions were detectable. The prevalence of the disease was quite low, which, together with the low morbidity observed in the lab, suggest DN is unlikely to be the direct cause of mortality outbreaks in France.

First description of a mortality event in adult Pacific oysters in Italy associated with infection by a Tenacibaculum soleae strain.

Summer mortality episodes in adult Pacific oysters have been described since the 1950s in various farming areas. Starting in 2012, a recrudescence of mortalities in commercial-sized oysters was first observed in France and then in Italy, with seasonality extension and translation later in the year. Moribund individuals collected during an event in Italy in December 2014 showed yellowish lesions of the mantle and adductor muscle. Histological examination revealed filamentous bacteria associated with necrotic areas. Quantitative PCRs targeting OsHV-1 and Vibrio aestuarianus detected only high loads of the pathogenic bacteria in tissues of symptomatic individuals. A lower diversity of the hemolymph microbiota was also evidenced in moribund individuals, with a predominance of Vibrio and Arcobacter species. A strain of Flavobacteriaceae was isolated from all the symptomatic individuals. Sequence analysis of the 16S rRNA gene identified the strain as Tenacibaculum soleae. When strain pathogenicity was tested by injection in adult individuals, it induced mortality rates of up to 45%, even in the absence of V. aestuarianus. As mortality occurred only 11 days post-infection, further investigation is needed to determine its effective virulence in natural conditions. This is the first description of a Tenacibaculum strain associated with bivalve mortalities.

Correcting symmetry imperfections in linear multipole traps.

Multipole radio-frequency (RF) traps are central to collisional experiments in cryogenic environments. They also offer possibilities to generate new type of ion crystals topologies and in particular the potential to create infinite 1D/2D structures: ion rings and ion tubes. However, multipole traps have also been shown to be very sensitive to geometrical misalignment of the trap rods, leading to additional local trapping minima. The present work proposes a method to correct non-ideal potentials, by modifying the applied RF amplitudes for each trap rod. This approach is discussed for the octupole trap, leading to the restitution of the ideal Mexican-Hat-like pseudo-potential, expected in multipole traps. The goodness of the compensation method is quantified in terms of the choice of the diagnosis area, the residual trapping potential variations, the required adaptation of the applied RF voltage amplitudes, and the impact on the trapped ion structures. Experimental implementation for macroscopic multipole traps is also discussed, in order to propose a diagnostic method with respect to the resolution and stability of the trap drive. Using the proposed compensation technique, we discuss the feasibility of generating a homogeneous ion ring crystal, which is a measure of quality for the obtained potential well.

Complete genome sequence of Ostreid herpesvirus type 1 µVar isolated during mortality events in the Pacific oyster Crassostrea gigas in France and Ireland.

Infections with Ostreid herpesvirus 1 (OsHV-1) microvariants in young Pacific oysters are associated with massive mortality events and significant economic losses. Previous studies, focusing on few regions of the genome, have revealed the genomic diversity of these genotypes with respect to the reference type. We used a NGS process to sequence the whole genome of the OsHV-1 µVar in infected individuals, collected during mortality events in France and Ireland. The final genome length of OsHV-1 µVar was approximately 205kbp, shorter than the reference genotype and the overall genome organisation resembled herpes simplex viruses. 94.4% similarity was observed with the OsHV-1 reference genotype. Large indels, including five deletions and three insertions were found to induce the loss and the addition of several ORFs, summed with codon substitutions in 64% of genes shared with the reference type. This diversity raises the question of the exact origin and evolution of OsHV-1 µVar.

Ostreid herpesvirus type 1 genomic diversity in wild populations of Pacific oyster Crassostrea gigas from Italian coasts.

Ostreid herpesvirus 1 (OsHV-1) is a significant pathogen affecting the young Pacific oyster Crassostrea gigas, worldwide. A new variant, OsHV-1 µVar, has been associated with recurrent mortality events in Europe since 2008. Epidemiological data collection is key for global risk assessment; however little is known about health status and genotypes present in European wild oyster beds. Most studies to date have involved only cultivated individuals during mortality events, and reported low genotype diversity. With this study, conducted along the Italian coasts, we investigated for the first time the presence of OsHV-1 in European natural oyster beds. Analysis of three genomic regions revealed the presence of at least nine different genotypes, including two variants close to the OsHV-1 reference, known since the early 1990s but with no European record reported since 2010, and highlights relevant genotype diversity in natural environment. Phylogenetic analysis distinguished two distinct clusters and geographical distribution of genotypes, with the exception of a variant very closely related to the µVar, which appeared the single genotype present in all the Adriatic sites. Interestingly, these wild symptom free populations could represent, in Europe, an accessible alternative to the import of OsHV-1-resistant oyster strains from the East Pacific, the native area of C. gigas, avoiding the high-risk of non-native marine species and new pathogen introductions.

Comparison of two real-time PCR methods for detection of ostreid herpesvirus 1 in the Pacific oyster Crassostrea gigas.

The real-time polymerase chain reaction (PCR) is considered to be a suitable tool for nucleic acid quantitation because it is accurate, rapid and reliable. The reference protocol for quantitation of ostreid herpesvirus 1 in Pacific oysters Crassostrea gigas is based on a Sybr(®) Green real-time PCR developed by the IFREMER laboratory. The Frank Duncombe Departmental Laboratory has developed an alternative protocol based on TaqMan(®) chemistry (alternative technique). The quantitation limits were 1000 and 18UG/mg of tissues for the reference method and alternative protocols, respectively, and the latter protocol has a detection limit of 6UG/mg of tissues. The aim of this study was to compare the two protocols using DNA samples obtained from 210 spat. The kappa index (0.41) indicated a moderate concordance between the protocols, according to the measures of Landis and Koch. All samples that were positive by the reference protocol were also positive by the alternative protocol. Of the 76 samples that were negative by the reference protocol, 49 were positives by the alternative protocol. In conclusion, the alternative protocol is an improvement of the reference protocol in terms of sensitivity, specificity and rapidity (<3h).

Terahertz frequency standard based on three-photon coherent population trapping.

A scheme for a terahertz frequency standard based on three-photon coherent population trapping in stored ions is proposed. Assuming the propagation directions of the three lasers obey the phase matching condition, we show that stability of few 10(-14) at 1 s can be reached with a precision limited by power broadening to 10(-11) in the less favorable case. The referenced terahertz signal can be propagated over long distances, the useful information being carried by the relative frequency of the three optical photons.

Magnetic resonance imaging of a brain abscess in a 10-month-old filly.

The purpose of this paper was to correlate the magnetic resonance imaging (MRI) characteristics of a mature brain abscess in a horse with histopathologic alterations of brain tissue. Eight months after the onset of clinical signs, MRI of the brain of a 10-month-old filly was performed. A large space-occupying lesion in the right cerebral hemisphere was identified. This space-occupying lesion was delineated by a thick and well-defined capsule that was isointense to brain parenchyma on the T1-weighted images and with a markedly hypointense on the T2-weighted images. The identification of such a capsule is highly diagnostic of a mature brain abscess. The lesion seen on MR images was confirmed at necropsy where a large abscess of the right hemisphere was observed. Streptococcus zooepidemicus and Pseudomonas aeruginosa were isolated from the abscess. Based on histopathologic examination, the signal characteristics of the capsule on T1-weighted and T2-weighted images were found to be due to the presence of numerous hemosiderin-laden macrophages. These results are in agreement with previous studies on human patients. This report confirms the value of MRI in the diagnosis of equine brain diseases.

Design and performance of a few-kilohertz-linewidth dye laserstabilized by reflection in an optical resonator.

Ultrahigh-resolution spectroscopy is limited by the spectral quality of lasers. We describe a servo system that stabilizes a dye laser by reflection in an optical resonator, which provides an error signal. We analyze the error signal to determine the spectral performance of the dye laser, which is shown to have a Gaussian-shaped spectrum with a width of less than 4 kHz.