Extended direct lysis method for virus detection on berries including droplet digital RT-PCR or real time RT-PCR with reduced influence from inhibitors

Detection of viruses on berries is a challenging task, often hampered by the presence of RT-qPCR inhibiting substances from berry juice. A direct extraction method for virus detection (murine norovirus and GA phage) on frozen raspberries was previously published. We expanded (different types of berries and viruses) and improved the method using MobiSpin S400 columns that filter nucleic acids based on size-exclusion chromatography. While no inhibition was detected in filtered RNA, unfiltered RNA needed from 1:2 to more than 1:8 dilution in order to remove inhibition. The modified method gave recoveries of bovine norovirus around 40.8 ± 4.5% (40.0 ± 7.0%), 48.0 ± 26.0% (50.5 ± 7.8%), 28.3 ± 2.6% (45.8 ± 6.6%) from frozen (fresh) raspberries, strawberries and blueberries, respectively. For the same samples, recoveries of hepatitis A virus were 34.0 ± 5.9% (34.0 ± 6.0%), 40.0 ± 13.3% (34.2 ± 10.5%) and 23.0 ± 6.8% (31.5 ± 7.9%). For adenovirus40 (DNA virus), recoveries were 21.2 ± 8.6%, 16.0 ± 3.2% and 5.7 ± 0.2% from fresh raspberries, strawberries and blueberries respectively and column filtration did not add any improved effect. The modified method is effective and timesaving for detection of viral RNA from both fresh and frozen berries. As an emerging detection and direct quantification method, droplet digital RT-PCR was compared to RT-qPCR and was much less influenced by inhibitors when detecting mengovirus in unfiltered RNA from berries. However, for low levels of pure RNA, RT-qPCR showed slightly higher sensitivity and more stable results

Spatio-temporal dynamics of virus and bacteria removal in dual-media contact-filtration for drinking water

Microorganism removal efficiencies in deep bed filters vary with time and depth in the filter bed as the filter collects particles. Improved knowledge of such dynamics is relevant for the design, operation and microbial risk assessment of filtration processes for drinking water treatment. Here we report on a highresolution spatio-temporal characterization of virus and bacteria removal in a pilot-scale dual-media filter, operated in contact-filtration mode. Microorganisms investigated were bacteriophage Salmonella typhimurium 28B (plaque assay, n ¼ 154)), fRNA phage MS2 (plaque assay/RT-qPCR, n ¼ 87) and E. coli (Colilert-18, n ¼ 73). Microscopic and macroscopic filtration models were used to investigate and characterize the removal dynamics. Results show that ripening/breakthrough fronts for turbidity, viruses and E. coli migrated in a wavelike manner across the depth of the filter. Virus removal improved continuously throughout the filter cycle and viruses broke through almost simultaneously with turbidity. Ripening for E. coli took longer than ripening for turbidity, but the bacteria broke through before turbidity breakthrough. Instantaneous log-removal peaked at 3.2, 3.0 and 4.5 for 28B, MS2 and E. coli, respectively. However, true average logremoval during the period of stable effluent turbidity was significantly lower at 2.5, 2.3 and 3.6, respectively. Peak observed filter coefficients l were higher than predicted by ideal filtration theory. This study demonstrates the importance of carefully designed sampling regimes when characterizing microorganism removal efficiencies of deep bed filters.Research Council of Norway (grant no. 226750/O30) and Norconsult AS consultancy firm14 Página

Bluetongue: a historical and epidemiological perspective with the emphasis on South Africa

Bluetongue (BT) is a non-contagious, infectious, arthropod transmitted viral disease of domestic and wild ruminants that is caused by the bluetongue virus (BTV), the prototype member of the Orbivirus genus in the family Reoviridae. Bluetongue was first described in South Africa, where it has probably been endemic in wild ruminants since antiquity. Since its discovery BT has had a major impact on sheep breeders in the country and has therefore been a key focus of research at the Onderstepoort Veterinary Research Institute in Pretoria, South Africa. Several key discoveries were made at this Institute, including the demonstration that the aetiological agent of BT was a dsRNA virus that is transmitted by Culicoides midges and that multiple BTV serotypes circulate in nature. It is currently recognized that BT is endemic throughout most of South Africa and 22 of the 26 known serotypes have been detected in the region. Multiple serotypes circulate each vector season with the occurrence of different serotypes depending largely on herd-immunity. Indigenous sheep breeds, cattle and wild ruminants are frequently infected but rarely demonstrate clinical signs, whereas improved European sheep breeds are most susceptible. The immunization of susceptible sheep remains the most effective and practical control measure against BT. In order to protect sheep against multiple circulating serotypes, three pentavalent attenuated vaccines have been developed. Despite the proven efficacy of these vaccines in protecting sheep against the disease, several disadvantages are associated with their use in the field

Microbiological food safety of seaweeds

The use of seaweeds in the human diet has a long history in Asia and has now been increasing also in the western world. Concurrent with this trend, there is a corresponding increase in cultivation and harvesting for commercial production. Edible seaweed is a heterogenous product category including species within the green, red, and brown macroalgae. Moreover, the species are utilized on their own or in combinatorial food products, eaten fresh or processed by a variety of technologies. The present review summarizes available literature with respect to microbiological food safety and quality of seaweed food products, including processing and other factors controlling these parameters, and emerging trends to improve on the safety, utilization, quality, and storability of seaweeds. The over- or misuse of antimicrobials and the concurrent development of antimicrobial resistance (AMR) in bacteria is a current worldwide health concern. The role of seaweeds in the development of AMR and the spread of antimicrobial resistance genes is an underexplored field of research and is discussed in that context. Legislation and guidelines relevant to edible seaweed are also discussed.publishedVersio

Temperature Dependent Depuration of Norovirus GII and Tulane Virus from Oysters (Crassostrea gigas)

Raw oysters are considered a culinary delicacy but are frequently the culprit in food-borne norovirus (NoV) infections. As commercial depuration procedures are currently unable to efficiently eliminate NoV from oysters, an optimisation of the process should be considered. This study addresses the ability of elevated water temperatures to enhance the elimination of NoV and Tulane virus (TuV) from Pacific oysters (Crassostrea gigas). Both viruses were experimentally bioaccumulated in oysters, which were thereafter depurated at 12 °C and 17 °C for 4 weeks. Infectious TuV and viral RNA were monitored weekly for 28 days by TCID50 and (PMAxx-) RT-qPCR, respectively. TuV RNA was more persistent than NoV and decreased by  1.0 log10 at this time. For RT-qPCR there was no detectable benefit of elevated water temperatures or PMAxx for either virus (p > 0.05). TuV TCID50 decreased steadily, and reductions were significantly different between the two temperatures (p  3.0 log10 were observed at 17 °C, while at 12 °C reductions did not exceed 1.9 log10. The length of depuration also had an influence on virus numbers. TuV reductions increased from  4.0 log10 after 4 weeks. This implies that an extension of the depuration period to more than seven days, possibly in combination with elevated water temperatures, may be beneficial for the inactivation and removal of viral pathogens.publishedVersio

Effectiveness of environmental surveillance of SARS-CoV-2 as an early warning system during the first year of the COVID-19 pandemic: a systematic review

publishedVersio