Nervous Necrosis Virus Replicates Following the Embryo Development and Dual Infection with Iridovirus at Juvenile Stage in Grouper

Infection of virus (such as nodavirus and iridovirus) and bacteria (such as Vibrio anguillarum) in farmed grouper has been widely reported and caused large economic losses to Taiwanese fish aquaculture industry since 1979. The multiplex assay was used to detect dual viral infection and showed that only nervous necrosis virus (NNV) can be detected till the end of experiments (100% mortality) once it appeared. In addition, iridovirus can be detected in a certain period of rearing. The results of real-time PCR and in situ PCR indicated that NNV, in fact, was not on the surface of the eggs but present in the embryo, which can continue to replicate during the embryo development. The virus may be vertically transmitted by packing into eggs during egg development (formation) or delivering into eggs by sperm during fertilization. The ozone treatment of eggs may fail to remove the virus, so a new strategy to prevent NNV is needed

Robust estimation of bacterial cell count from optical density

Optical density (OD) is widely used to estimate the density of cells in liquid culture, but cannot be compared between instruments without a standardized calibration protocol and is challenging to relate to actual cell count. We address this with an interlaboratory study comparing three simple, low-cost, and highly accessible OD calibration protocols across 244 laboratories, applied to eight strains of constitutive GFP-expressing E. coli. Based on our results, we recommend calibrating OD to estimated cell count using serial dilution of silica microspheres, which produces highly precise calibration (95.5% of residuals <1.2-fold), is easily assessed for quality control, also assesses instrument effective linear range, and can be combined with fluorescence calibration to obtain units of Molecules of Equivalent Fluorescein (MEFL) per cell, allowing direct comparison and data fusion with flow cytometry measurements: in our study, fluorescence per cell measurements showed only a 1.07-fold mean difference between plate reader and flow cytometry data

An automated microfluidic chip system for detection of piscine nodavirus and characterization of its potential carrier in grouper farms.

Groupers of the Epinephelus spp. are an important aquaculture species of high economic value in the Asia Pacific region. They are susceptible to piscine nodavirus infection, which results in viral nervous necrosis disease. In this study, a rapid and sensitive automated microfluidic chip system was implemented for the detection of piscine nodavirus; this technology has the advantage of requiring small amounts of sample and has been developed and applied for managing grouper fish farms. Epidemiological investigations revealed an extremely high detection rate of piscine nodavirus (89% of fish samples) from 5 different locations in southern Taiwan. In addition, positive samples from the feces of fish-feeding birds indicated that the birds could be carrying the virus between fish farms. In the present study, we successfully introduced this advanced technology that combines engineering and biological approaches to aquaculture. In the future, we believe that this approach will improve fish farm management and aid in reducing the economic loss experienced by fish farmers due to widespread disease outbreaks

Symptomatic Resolution Among Chinese Patients With Schizophrenia and Associated Factors

The remission and resolution criteria for schizophrenia were defined by the Remission in Schizophrenia Working Group in 2005, using eight core items of the positive and negative symptoms scale for schizophrenia. Subsequent studies of Caucasians have reported similar remission/resolution rates of approximately one-third. However, the remission/resolution rate in Chinese patients has not previously been reported. The present study assessed symptom resolution rates and associated factors among medicated and clinically stable Chinese schizophrenia patients. Methods: Chinese patients with a diagnosis of schizophrenia were followed-up 1 month after their last psychiatric hospitalization. Cross-sectional clinical assessments for psychopathology, side effect profiles, quality of life, psychosocial function, and neurocognition tests were performed. Results: Thirty-three (36.7%) of a total of 90 patients met the resolution criteria. They had a significantly higher level of education and lower scores for positive symptoms, negative symptoms, and general psychopathology on the positive and negative symptoms scale; they had lower scores on the Udvalg for Kliniske Undersøgelser Side Effect Rating Scale, and Simpson Angus Scale; and higher scores on the Global Assessment of Functioning and Subjective Well-being under Neuroleptics scales, compared with patients who did not meet the resolution criteria. Multiple regression analyses controlling for age, sex, duration of illness, education, duration of index hospitalization, and antipsychotic dosage revealed that a higher Udvalg for Kliniske Undersøgelser Side Effect Rating Scale score was related to lower rate of symptom resolution. The patients treated with clozapine and combinations of first generation antipsychotics and second generation antipsychotics had more severe psychopathology and side effects and showed a significantly lower resolution rate than did patients treated with first generation antipsychotics or second generation antipsychotics alone. Conclusion: Consistent with studies of Caucasian patients, one-third of clinically stable Chinese patients met the resolution criteria, as well as having fewer general side effects, better global functioning and subjective well-being

Detection of potential nervous necrosis virus (NNV) carriers.

a<p>Birds commonly seen in grouper fish farms are <i>Egretta garzetta</i> (little egret), <i>Nycticorax nycticorax</i> (black-crowned night heron), and <i>Passer montanus</i> (tree sparrow).</p>b<p>Brine shrimps used as a feed for larvae.</p>c<p>Common water fleas.</p>d<p>(number of positive/number of negative); +, NNV positive; −, NNV negative.</p

Design of integrated microfluidic chip.

<p>A. Schematic diagram of the integrated microfluidic chip. The micro RT-PCR module, capillary electrophoresis (CE) module and buried optical fibers were integrated; (a), reaction chamber; (b), RT reagent reservoir; (c), PCR reagent reservoir; (d), sample reservoir; (e), CE sample reservoir; (f), buffer reservoir; (g), waste collection chamber. B. Photograph of the microfluidic chip. The 3 layers of the integrated chip are composed of a polymethylmethacrylate (PMMA) CE chip, a glass micro PCR chip, and Polydimethylsiloxane (PDMS) micropumps. The dimension of chip is 6.5×3.9×0.8 cm³. The CE channel is 100 µm in width ×30 µm in depth. The volumes of the PCR and reagent chambers are 11.25 µL.</p

Neural Network of Body Representation Differs between Transsexuals and Cissexuals

<div><p>Body image is the internal representation of an individual’s own physical appearance. Individuals with gender identity disorder (GID), commonly referred to as transsexuals (TXs), are unable to form a satisfactory body image due to the dissonance between their biological sex and gender identity. We reasoned that changes in the resting-state functional connectivity (rsFC) network would neurologically reflect such experiential incongruence in TXs. Using graph theory-based network analysis, we investigated the regional changes of the degree centrality of the rsFC network. The degree centrality is an index of the functional importance of a node in a neural network. We hypothesized that three key regions of the body representation network, i.e., the primary somatosensory cortex, the superior parietal lobule and the insula, would show a higher degree centrality in TXs. Twenty-three pre-treatment TXs (11 male-to-female and 12 female-to-male TXs) as one psychosocial group and 23 age-matched healthy cissexual control subjects (CISs, 11 males and 12 females) were recruited. Resting-state functional magnetic resonance imaging was performed, and binarized rsFC networks were constructed. The TXs demonstrated a significantly higher degree centrality in the bilateral superior parietal lobule and the primary somatosensory cortex. In addition, the connectivity between the right insula and the bilateral primary somatosensory cortices was negatively correlated with the selfness rating of their desired genders. These data indicate that the key components of body representation manifest in TXs as critical function hubs in the rsFC network. The negative association may imply a coping mechanism that dissociates bodily emotion from body image. The changes in the functional connectome may serve as representational markers for the dysphoric bodily self of TXs.</p></div

Optimization of the multiplex RT-PCR system.

<p>The microfluidic RT-PCR system is comprised of a microfluidic control module and a micro temperature control module <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0036183#pone.0036183-Lien1" target="_blank">[18]</a>. The current design of microfluidic system is capable for rapid detection of four target genes (two viruses, one bacteria and one host immune response gene). The cDNA of samples can be uniformly distributed into the subsequent four PCR reaction chambers to detect four genes simultaneously. <b>A</b>. 100 ng of cDNA was amplified with each set of primers separately and with the mixture of the primers. Lane 2, Iridovirus; lane 3, <i>Vibrio anguillarum</i>; lane 4, Mx; lane 5, NNV (nodavirus); lane 1, multiprimer reactions simultaneously in a single tube. <b>B</b>. The electropherograms of the RT-PCR products from the single-tube reaction. The sizes of the DNA marker are 310 bp, 281 bp, 271 bp, 234 bp, 194 bp, 118 bp and 72 bp.</p