Rapid sample preparation and low-resource molecular detection of hepatopancreatic parvoviruses (HPV) by recombinase polymerase amplification lateral flow detection assay in shrimps (Fenneropenaeus merguiensis)

Background Viral diseases are a major problem in shrimp aquaculture facilities as these diseases reduce growth rates, which inevitably lead to production and profit losses. Hepatopancreatic parvoviruses (HPV) are common diseases in shrimp that appear to be associated with high or low levels of replication in specific genetic lineages. Selective breeding may result in resistance to HPV and improved body traits such as body weight, meat yield and shrimp colour, facilitating shrimp farming. HPV virus titre is commonly determined by quantitative PCR (qPCR), which is a time-consuming method requiring laboratory equipment unsuitable for field implementation. The aim of this study was to develop a simple, robust, rapid and reliable method to detect HPV in low-resource environments. Methods We developed a rapid shrimp HPV test that uses (1) a simple three-step sample preparation protocol, followed by (2) isothermal recombinase polymerase amplification (RPA) and lateral flow strip detection (LFD). Analytical sensitivity testing was performed in a background banana shrimp sample matrix, and retrospective testing of Fenneropenaeus merguiensis hepatopancreas tissues (n = 33) with known qPCR viral titres was used to determine diagnostic sensitivity and specificity. Results The rapid shrimp HPV test could detect as little as 35 genome-equivalent copies per reaction in homogenized F. merguiensis banana shrimp. Retrospective testing of stored tissues (n = 33) indicated 100% diagnostic sensitivity (95% confidence interval, CI: 86–100%) and 100% specificity (95% CI: 66–100%) for detection of HPV. Conclusion The rapid shrimp HPV test could be completed in only 40 minutes, and required only homogenization pestles, some pipettors, and a small heating block for single temperature incubation at 39°C. Critically, our procedure eliminated the time-consuming purification of nucleic acids from samples and when combined with RPA-LFD offers a user-friendly HPV detection format that can potentially be performed on-site. Our approach represents a major step forward in the development of a simple and sensitive end-point method for quick determination of unfavourable HPV virus numbers in shrimp, and has great potential to advance on-site management of shrimps in aquaculture

Dolly Bear Shop / Mohd Nor Hafizie Othman ... [et al.]

After we have a big discussion, finally we decide to choose the type of business is in state of a partnership business because it is a starting point to establish a business.Our company name is 'DOLLY BEAR SHOP' where the chosen based on our product, because we planned to sell the kind of soft toys which is dolls and bears. Since our business is partnership, so our ownership structure revolves among shareholders of members.This business owned by 5 members of us and each of us play an important role to ensure this business achieving the target and mission.Our director is known as General Manager and followed by 4 managers which is Administrative Manager, Marketing Manager, Operation Manager and Financial Manager who is responsible to supervise and to make sure the business run smoothly with strong financial condition so that we can maintain our business and make our shop famous

Abundance and distribution of sperm whales in the Canary Islands : can sperm whales in the Archipelago sustain the current level of ship-strike mortalities?

Funding was provided through an agreement between the Canary Islands Government and the Spanish Ministries of the Environment and Defence. Additional survey effort on the Amanay, Banquete and Concepción seamounts was funded by the Fundación Biodiversidad-MAGRAMA via the LIFE-INDEMARES project.Sperm whales are present in the Canary Islands year-round, suggesting that the archipelago is an important area for this species in the North Atlantic. However, the area experiences one of the highest reported rates of sperm whale ship-strike in the world. Here we investigate if the number of sperm whales found in the archipelago can sustain the current rate of ship-strike mortality. The results of this study may also have implications for offshore areas where concentrations of sperm whales may coincide with high densities of ship traffic, but where ship-strikes may be undocumented. The absolute abundance of sperm whales in an area of 52933 km2, covering the territorial waters of the Canary Islands, was estimated from 2668 km of acoustic line-transect survey using Distance sampling analysis. Data on sperm whale diving and acoustic behaviour, obtained from bio-logging, were used to calculate g(0) = 0.92, this is less than one because of occasional extended periods when whales do not echolocate. This resulted in an absolute abundance estimate of 224 sperm whales (95% log-normal CI 120-418) within the survey area. The recruitment capability of this number of whales, some 2.5 whales per year, is likely to be exceeded by the current ship-strike mortality rate. Furthermore, we found areas of higher whale density within the archipelago, many coincident with those previously described, suggesting that these are important habitats for females and immature animals inhabiting the archipelago. Some of these areas are crossed by active shipping lanes increasing the risk of ship-strikes. Given the philopatry in female sperm whales, replacement of impacted whales might be limited. Therefore, the application of mitigation measures to reduce the ship-strike mortality rate seems essential for the conservation of sperm whales in the Canary Islands.Publisher PDFPeer reviewe

Correction: Abundance and Distribution of Sperm Whales in the Canary Islands: Can Sperm Whales in the Archipelago Sustain the Current Level of Ship-Strike Mortalities?

[This corrects the article DOI: 10.1371/journal.pone.0150660.]

Metformin inhibits cell cycle progression of B-cell chronic lymphocytic leukemia cells.

B-cell chronic lymphocytic leukemia (CLL) was believed to result from clonal accumulation of resting apoptosis-resistant malignant B lymphocytes. However, it became increasingly clear that CLL cells undergo, during their life, iterative cycles of re-activation and subsequent clonal expansion. Drugs interfering with CLL cell cycle entry would be greatly beneficial in the treatment of this disease. 1, 1-Dimethylbiguanide hydrochloride (metformin), the most widely prescribed oral hypoglycemic agent, inexpensive and well tolerated, has recently received increased attention for its potential antitumor activity. We wondered whether metformin has apoptotic and anti-proliferative activity on leukemic cells derived from CLL patients. Metformin was administered in vitro either to quiescent cells or during CLL cell activation stimuli, provided by classical co-culturing with CD40L-expressing fibroblasts. At doses that were totally ineffective on normal lymphocytes, metformin induced apoptosis of quiescent CLL cells and inhibition of cell cycle entry when CLL were stimulated by CD40-CD40L ligation. This cytostatic effect was accompanied by decreased expression of survival- and proliferation-associated proteins, inhibition of signaling pathways involved in CLL disease progression and decreased intracellular glucose available for glycolysis. In drug combination experiments, metformin lowered the apoptotic threshold and potentiated the cytotoxic effects of classical and novel antitumor molecules. Our results indicate that, while CLL cells after stimulation are in the process of building their full survival and cycling armamentarium, the presence of metformin affects this process

Probability histograms.

<p>Probability histograms of the duration of echolocating and not echolocating phases (in 1 minute-bins) for the seven sperm whales tagged off the Azores. The echolocation phase represents the time elapsed from the start to the end of ‘usual’ clicking in a foraging dive; the silent phase, i.e. when the whale was not echolocating, is the sum of the silent descent, silent ascent and the time spent at the surface between dives (IDI). The simulation model was run with parameters chosen from these echolocating/not echolocating distributions.</p

Survey area.

<p>Survey area indicating the names of the islands and seamounts, and showing selected isobaths, the survey blocks (black lines) and the designed transects (magenta lines).</p

Sperm whale survey effort and detections overlaid on map of marine traffic density.

<p>Acoustically surveyed tracks are shown as green lines, while tracks in red show when sperm whales were acoustically detected. Marine traffic data were obtained from AIS data and coloured by traffic density (given as AIS signals/km²/year).</p

Information on the sperm whale DTag recordings.

<p>Whale codes are generated by combining the three-digit Julian day and the deployment order of the tag in that day (as a letter). A complete dive cycle comprises a foraging dive and the following inter-dive interval. Only complete dive cycles were used here, and the first dive cycle after tagging was excluded. Foraging dives were divided into an echolocation period (when animals were producing 'usual' clicks [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0150660#pone.0150660.ref030" target="_blank">30</a>]) and periods of silent descent and ascent (between the surface and the depth at which animals started and stopped clicking). Durations are given as the median of each period (range).</p

Histogram and detection function.

<p>Histogram of perpendicular distances to sperm whale acoustic detections and the fitted detection function. Data were grouped into 600 m bins for the histogram, although exact distances were used to fit the detection function. Data were truncated at 9 km.</p