Suchergebnisse

In last five years, the Africa has faced two outbreaks of Zaire ebolavirus. These outbreaks have been the largest so far, and latest outbreak is still ongoing and affecting the Democratic Republic of the Congo. We tested in parallel three different Zaire ebolavirus (EBOV) realtime RT-PCRs targeting the nucleoprotein gene (EBOV NP-RT-qPCRs) described by Trombley et al. (2010), Huang et al. (2012) and Weidmann et al. (2004). These assays are used regularly in diagnostic laboratories. The limit of detection (LOD), intra-assay repeatability using different matrixes, sensitivity and specificity were determined. In addition, the primers and probes were aligned with the sequences available in ongoing and past outbreaks in order to check the mismatches. The specificity of all three EBOV NP-RT-qPCRs were excellent (100%), and LODs were under or 10 copies per PCR reaction. Intra-assay repeatability was good in all assays, however the Ct-values were bit higher using the EDTA-blood based matrix. All of the primers and probes in EBOV NP-RTqPCR assays have one or more mismatches in the probes and primers when the 2267 Zaire EBOV NP sequences, including strains Ituri from DRC outbreak (year 2018), was aligned. The EBOV strain of Bikoro (year 2018) circulating in DRC was 100% match in Trombley and Weidmann assay, but had one mismatch in Huang assay.

Threshold infection level is the pathogen load of the test animals measured before the appearance of clinical signs and mortality. This study aims to establish the threshold infection levels of WSSV in different weight ranges of Penaeus vannamei using qPCR. Artificial infection experiments were conducted using four weight ranges (3–5 g, 7–8 g, 15–18 g, and 22–25 g). The LD 50 of the different weight ranges of shrimps were achieved at viral dilution of 10 -6 and 10 -5 after 216–240 hpi, and the viral loads of these inoculums have a range of 10 5 –10 6 WSSV DNA copies/g. The viral loads of the samples in the timecourse infection experiments when the mortalities started was determined at 10 9 WSSV DNA copies/g, while for the survivors was at 10 6 WSSV DNA copies/g. The threshold infection level of WSSV in shrimp was determined at 10 7 to 10 8 WSSV DNA copies/g. It was also found out that the threshold infection level was not weight dependent. ; We are very grateful to the Government of Japan-Trust Fund (study code: FH-07- F2015T) and SEAFDEC/AQD for funding this work. We thank Mr. Ivan Yaptangco of IJY Shrimp Farm for providing the experimental animals and the staff of Fish Health Section Diagnostic Services for the technical support.

Threshold infection level is the pathogen load of the test animals measured before the appearance of clinical signs and mortality. This study aims to establish the threshold infection levels of WSSV in different weight ranges of Penaeus vannamei using qPCR. Artificial infection experiments were conducted using four weight ranges (3–5 g, 7–8 g, 15–18 g, and 22–25 g). The LD 50 of the different weight ranges of shrimps were achieved at viral dilution of 10 -6 and 10 -5 after 216–240 hpi, and the viral loads of these inoculums have a range of 10 5 –10 6 WSSV DNA copies/g. The viral loads of the samples in the timecourse infection experiments when the mortalities started was determined at 10 9 WSSV DNA copies/g, while for the survivors was at 10 6 WSSV DNA copies/g. The threshold infection level of WSSV in shrimp was determined at 10 7 to 10 8 WSSV DNA copies/g. It was also found out that the threshold infection level was not weight dependent. ; We are very grateful to the Government of Japan-Trust Fund (study code: FH-07- F2015T) and SEAFDEC/AQD for funding this work. We thank Mr. Ivan Yaptangco of IJY Shrimp Farm for providing the experimental animals and the staff of Fish Health Section Diagnostic Services for the technical support.

What is the impact of the recent Data Access and Research Transparency (DA-RT) initiative and the Journal Editors Transparency Statement (JETS) on scholars working with qualitative data? Analysis finds DA-RT insufficiently sensitive to the needs of qualitative data and focuses on four inter-related reasons why DA-RT needs to be revised before being widely adopted by political science journals: (1) space constraints that hinder full journal presentation of the analysis of qualitative data; (2) ethical concerns about protecting human subjects, and the time needed to prepare such data before publicly sharing them; (3) costs of data collection and the right of first usage; and (4) a potentially chilling effect of DA-RT on certain types of research topics. Analysis of the author's own journey from econometric and survey analysis to narrative interviews with people in vulnerable situations, facing moral dilemmas, illustrates why DA-RT needs additional safeguards for qualitative data and methods. Given the increasing importance of qualitative data, and its ability to lend insight into critical political topics, the author argues that implementing the current version of the DA-RT initiative could hinder political science's ability to address important political questions. Thus DA-RT must be modified to address the special needs of qualitative data.

Biological indexing is the method generally recognized for the certification of propagative grapevines in many countries, and it is mandatory in the European Union. It consists of the evaluation of the plant material after grafting on indicators that are inspected for symptom development. This is a lengthy process that requires well-trained workers, testing field, etc. Alternative diagnostic methods such as serology and RT-qPCR have been discarded for certification because of their intrinsic drawbacks. In turn, high-throughput sequencing (HTS) of plant RNA has been proposed as a plausible alternative to bioassay, but before it is accepted, different aspects of this process must be evaluated. We have compared the HTS of small RNAs with bioassays and other diagnostic methods from a set of 40 grapevine plants submitted for certification. The results allowed the authors the identification of numerous grapevine viruses in the samples, as well as different variants. Besides, relationships between symptom expression and viromes were investigated, in particular leafroll-associated viruses. We compared HTS results using analytical and bioinformatics approaches in order to define minimum acceptable quality standards for certification schemes, resulting in a pipeline proposal. Finally, the comparison between HTS and bioassay resulted favorable for the former in terms of reliability, cost, and timing.

Hepatitis E virus (HEV) is an important enteric agent that can circulate in swine; it is excreted in manure, and of zoonotic interest. The present study investigated, by RT-qPCR, the circulation of HEV in swine manure from different types of pig farms (maternity, nursery, and grow-finish farms) in Santa Catarina State, the major pig production area of Brazil, and also evaluated the HEV removal efficiency of psychrophilic anaerobic biodigesters (PABs). While HEV was consistently detected in manure from grow-finish pig farms (>4 log HEV genome copies (GC) L(−1)), the virus was not detected in manure from maternity and nursery farms. These findings suggest a potential high biosafety status during primary-swine production, with a subsequent contamination in grow-finish production. The anaerobic biodigestion process reduced more than 2 log(10) HEV GC in the processed swine manure. However, the virus concentration in final effluent remained high, with an average value of 3.85 log(10) HEV GC L(−1). Consequently, our results demonstrate that PABs can be a robust tool for effective inactivation of HEV, while reinforcing the need for sanitary surveillance and legislation of swine manure-derived biofertilizers, to avoid the spread of zoonotic enteric pathogens such as HEV.