Rabbit haemorrhagic disease virus Lagovirus europaeus/GI.1d strain: genome sequencing, in vivo virus replication kinetics, and viral dose effect

Background Rabbit haemorrhagic disease virus Lagovirus europaeus/GI.1d variant (GI.1d/RHDV) was identified in 1990 in France, and until the emergence of the new genotype GI.2, it was the main variant circulating in the country. The early stages of RHDV infection have been described in a few studies of rabbits experimentally infected with earlier strains, but no information was given on the minimum infective dose. We report the genomic and phenotypic characterisation of a GI.1d/RHDV strain collected in 2000 in France (GI.1d/00–21). Results We performed in vivo assays in rabbits to study virus replication kinetics in several tissues at the early stage of infection, and to estimate the minimum infective dose. Four tested doses, negligible (10− 1 viral genome copies), low (104), high (107) and very high (1011) were quantified using a method combining density gradient centrifugation of the viral particles and an RT-qPCR technique developed to quantify genomic RNA (gRNA). The GI.1d/00–21 genome showed the same genomic organisation as other lagoviruses; however, a substitution in the 5′ untranslated region and a change in the potential p23/2C-like helicase cleavage site were observed. We showed that the liver of one of the two rabbits inoculated via the oral route was infected at 16 h post-infection and all tissues at 39 h post-infection. GI.1d/00–21 induced classical RHD signs (depression) and lesions (haemorrhage and splenomegaly). Although infective dose estimation should be interpreted with caution, the minimum infective dose that infected an inoculated rabbit was lower or equal to 104 gRNA copies, whereas between 104 and 107 gRNA copies were required to also induce mortality. Conclusions These results provide a better understanding of GI.1d/RHDV infection in rabbits. The genome analysis showed a newly observed mutation in the 5′ untranslated region of a lagovirus, whose role remains unknown. The phenotypic analysis showed that the pathogenicity of GI.1d/00–21 and the replication kinetics in infected organs were close to those reported for the original GI.1 strains, and could not alone explain the observed selective advantage of the GI.1d strains. Determining the minimum dose of viral particles required to cause mortality in rabbits is an important input for in vivo studies.

Transmission, infection, and pathogenesis

Transmission is a key process for parasites. Different routes (e.g. faecal–oral) and modes (e.g. by aerosols or vectors) exist. A major context is vertical (to offspring) or horizontal (all other) transmission. All components of the transmission process evolve. Successful transmission includes the infection of a new host. Macroparasites typically infect as individuals, but microparasites need an infective dose. Doses vary enormously among parasites. Various models describe variation in infective dose. Process-based models assume random colonization, co-operative parasite manipulation, or are focused on early dynamics. With the processes of pathogenesis (e.g. tissue destruction, reducing host capacities), damage to the host emerges. Virulence factors are important mediators of parasite success and often involved in host manipulation and pathogenesis, including immunopathology.

shRNA targeting nonstructural protein inhibits the replication of severe fever with thrombocytopenia syndrome virus

Aim: To explore whether shRNA targeting nonstructural protein (NSs) of severe fever with thrombocytopenia syndrome virus (SFTSV) could inhibit SFTSV replication in Vero cells. Materials & methods: SFTSV used in this experiment was propagated in Vero cells and stored at -20°C. shRNA plasmid against NSs of SFTSV was transfected to Vero cells and infected with SFTSV, after which western blotting and tissue culture infective dose (TCID50) were used to measure the virus titers. Results: shRNA against NSs protein decreased the expression of NSs and inhibited the replication of SFTSV. Conclusion: The constructed SFTSV NSs-shRNA plasmid could inhibit the replication of SFTSV. It was concluded that SFTSV NSs-shRNA could inhibit virus replication for at least 72 h. shRNA-mediated antiviral effects were dose-dependent.

Microbial dynamic and growth potential of selected pathogens in Ethiopian traditional fermented beverages

Purpose The patterns of microbial succession and the associated physicochemical changes in the course of beverage fermentation determine the safety status of the final product against foodborne pathogens. In this study, the microbial dynamics during fermentation of three Ethiopian traditional fermented beverages (namely, borde, tej, and grawa) and the growth potential of selected foodborne pathogens in ready-to-consume beverages were assessed. Methods The raw materials used for lab-scale fermentation of the beverages were bought from open markets of Jimma and Anfilo towns. During fermentation, samples were drawn every 6 h (borde fermentation) and 12 h (grawa and tej fermentation). The dominant microbes of the fermentation phases were determined following standard microbiological methods. The growth potential of Escherichia coli, Salmonella typhimurium, Staphylococcus aureus, Listeria monocytogenes, and Candida albicans in the ready-to-consume beverages were assessed by microbial enumeration over defined storage period. Result Early fermentation period of all beverages was dominated by aerobic mesophilic bacteria, staphylococci, and Enterobacteriaceae with highest mean counts (Log CFU/ml) of 6.42 ± 0.10, 5.44 ± 0.08, and 5.40 ± 0.11, respectively. At the end of fermentation, yeast counts (Log CFU/ml) dominated in tej (9.41 ± 0.06) and grawa (7.88 ± 0.02) samples, while lactic acid bacteria dominated in borde sample (7.33 ± 0.07). During fermentation, pH dropped for borde (4.58 ± 0.03 to 4.22 ± 0.01), and grawa (4.18 ± 0.10 to 3.62 ± 0.02), but increased for tej (5.26 ± 0.01 to 5.50 ± 0.03) during the first 24 h, though it dropped later down to 3.81 ± 0.02 at 144th h. All reference pathogens were unable to reach infective dose in grawa and tej samples. However, borde sample supported their growth to infective dose within 24 h. Thus, grawa and tej beverages had the capability of inhibiting growth of pathogens while borde needs basic safety control measures during preparation and storage. Conclusion With further safety evaluation of the products, the production processes of the three beverages could be scaled up for commercial purposes using defined starter cultures originated from the same beverages. However, the safety status of borde calls for further evaluation for alternative shelf-life extension mechanisms including the introduction of organic preservatives from local products such as medicinal plants.

Probability of infectious disease in humans during epidemic

Popular SIR models and their modifications used to generate predictions about epidemics and, specifically, the COVID-19 pandemic, are inadequate. The aim of this study was to find the laws describing the probability of infection in a biological object. Using theoretical methods of research based on the probability theory, we constructed the laws describing the probability of infection in a human depending on the infective dose and considering the temporal characteristics of a given infection. The so-called generalized time-factor law, which factors in the time of onset and the duration of an infectious disease, was found to be the most general. Among its special cases are the law describing the probability of infection developing by some point in time t, depending on the infective dose, and the law that does not factor in the time of onset. The study produced a full list of quantitative characteristics of pathogen virulence. The laws described in the study help to solve practical tasks and should lie at the core of mathematical epidemiological modeling.

Review of Infective Dose, Routes of Transmission, and Outcome of COVID-19 Caused by the SARS-CoV-2 Virus: Comparison with Other Respiratory Viruses

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is pandemic. Prevention and control strategies require an improved understanding of SARS-CoV-2 dynamics. We did a rapid review of the literature on SARS-CoV-2 viral dynamics with a focus on infective dose. We sought comparisons of SARS-CoV-2 with other respiratory viruses including SARS-CoV-1 and MERS-CoV. We examined laboratory animal, and human studies. The literature on infective dose, transmission, and routes of exposure was limited specially in humans, and varying endpoints were used for measurement of infection. We propose the minimum infective dose of COVID-19 in humans, is higher than 100 particles, possibly slightly lower than the 700 particles estimated for H1N1 influenza. Despite variability in animal studies, there was some evidence that increased dose at exposure correlated with higher viral load clinically, and severer symptoms. Higher viral load measures did not reflect COVID-19 severity. Aerosol transmission seemed to raise the risk of more severe respiratory complications in animals. An accurate quantitative estimate of the infective dose of SARS-CoV-2 in humans is not currently feasible and needs further research. Further work is also required on the relationship between routes of transmission, infective dose, co-infection, and outcomes.

Aerosol Release by Healthy People during Speaking: Possible Contribution to the Transmission of SARS-CoV-2

Our research aimed to review the potential risk of infection by SARS-CoV-2. We used an excerpt of a data set generated in May 2020 for reviewing the SARS-CoV-2 prevention concept of orchestras, singers and actors. People were sampled for droplet release for one-hour activities using a Grimm spectrometer covering a spectrum of 1 to 32 µm diameter. We estimated the number of “quanta” in the exhaled liquid from viral concentrations of 106 to 1011/mL, based on the Human Infective Dose 50 of 218 viral particles. We employed the Wells–Riley equation to estimate the risk of infection in typical meeting rooms for a one-hour meeting of 2, 4 and 6 people observing a 2 m distance. The four participating adults released a mean of 1.28 nLm3 while breathing, 1.68 nL/m3 while speaking normally, and two adults released a mean of 4.44 nL/m3 while talking with a raised voice. The combination of 50% breathing, 45% talking normally and 5% speaking with a raised voice increased the risk of infection above 5% for a one-hour meeting of two people. The result is based on 6 quanta released, corresponding to an initial virus concentration of 1000/nL (109/mL) in the fluid of the upper respiratory tract. Our data confirm the importance of using facemasks in combination with other measures to prevent transmission of SARS-CoV-2 at the workplace.