Tritrichomonas foetus. Thirty-three US states have state rules on this disease and render it reportable due to potential huge economic losses to cattle industry. The various rules of different states generally mandate testing and culling T. foetus-positive bulls as well as prohibiting import of T. foetus-positive animals. Wyoming has enforced these rules for over 20 year beginning in 2000. From 2017 to 2019, 3 years in a row, not even one T. foetus-positive bull has been detected throughout the entire state among over ten thousand bulls tested annually. Wyoming is the first US state to achieve total control and eradication of bovine trichomonosis by testing and culling T. foetus-positive bulls.Bovine trichomonosis is caused by  Tritrichomonas foetus, which is closely related to a human trichomonad protozoan Trichomonas vaginalis. The parasite is transmitted by direct sexual contact. An infected bull may appear normal without clinical signs except a preputial discharge associated with small nodules on the preputial and penile membranes in the early stage of infection [T. foetus-positive bull is always a major, if not the solely source of infection for a herd of cows served with natural mating. In contrast, a female cattle experiences vaginitis, endometritis, early abortion and transient or even permanent infertility after infection [T. foetus-positive cattle herd [Bovine trichomonosis is a sexually transmitted disease in cattle. Its causative pathogen is nfection , 2. Nevenfection , 2. Thernfection . A cow otle herd .Tritrichomonas foetus and bovine trichomonosis it causes in cattle have been found or presume to be worldwide on all continents except the Antarctica [T. foetus-positive bulls as well as prohibiting import of positive ones. These rules aim to control and eventually eradicate bovine trichomonosis.tarctica –8. Bovintarctica . Recentltarctica –12. Due T. foetus-infected bulls and open cows; and (2) lower calf crop as a result of fewer calves born and lighter calf bodyweight.Accurately calculating the economic losses caused by bovine trichomonosis on both individual cattle producers and cattle industry as a whole is very challenging if not impossible. Economic losses caused by bovine trichomonosis mainly consist of diminished financial gains due to (1) financial costs of testing bulls and veterinary expenses, culling and replacing T. foetus test is $8.00/each for 1–2 samples or $6.00/each for ≥ 3 samples using culturing; $30.00/each for 1–9 samples or $25.00/each for ≥ 10 samples using PCR [Factors that affect the cost for testing bulls include, but are not limited to, farm size in numbers of cows, bull to cow ratio, distance to veterinarian service and numbers of test performed. An estimate for bull-test cost per cow was made based on a survey of veterinarians in Colorado State is showed in Table sing PCR . Using tT. foetus. The model showed that (1) 14–50% reduction in annual calf crop; (2) 5–12% reduction in sucking/growing period; (3) 4–10% reduction in the bodyweight of calf crop at weaning; (4) 4–10% reduction in monetary return per calf born and (5) 5–35% reduction in in monetary return per cow [In Wyoming it was estimated that a 20% reduction in calf crop due to bovine trichomonosis in a 100-cow herd in 2011 could cause $20,000 in annual losses . Further per cow .T. foetus-positive bulls as well as prohibiting import of T. foetus-positive bulls. At the same time the rule listed bovine trichomonosis a state notifiable disease. The disease had been known in the state since 1970s, and the WSVL performed only hundreds of tests on bovine samples for T. foetus annually prior to 2000. Since then the numbers of samples tested annually have exponentially increased to thousands. The author had served as the WSVL Parasitology Section head between 2008 and 2013. A few specific aspects are worth discussion for the Wyoming’s journey.Wyoming started its journey for control and eradication of bovine trichomonosis in the year 2000 when it started a state rule called “Chapter 15” that mandates testing all bulls grazing on open/public allotments or being traded or leased for reproduction and culling T. foetus-infected cattle herds. These were on producers , herds  and allotments  in neighborhood). An overall response rate was 23.4% (1288/5498). Risk factors identified and their odds ratio (OR) were: neighboring a positive herd(s) , grazing on public allotments , commingling with other herd  and elapsed time taken to fix a broken fence(s) a week or longer  [A statewide survey of all cattle producers in Wyoming was carried out in 2011 by the author and colleagues using a questionnaire. Twenty-five variables were included in the questionnaire for their possible association with the positive status among the .9–20.2) .In the USA, cattle producers are voluntary in performing AI among their cattle herds. Not a single US state has a decree that demands mandatory AI. In the above-mentioned questionnaire cattle producers were also surveyed for breeding methods in their cattle herds. Overwhelming 80.8% of cattle herds were by live bull service whereas only 2.1% was by AI in the year 2010. Furthermore, 56.1% of producers in Wyoming would not consider AI and only 36.7% would in the future . TherefoT. foetus from Wyoming positive bulls were cultured and their identity was investigated by sequencing partial small and large subunit rRNA, the internal transcribed spacer 1 and 2, and 5S rRNA fragments followed by a phylogenetic analysis. All 24 isolates had identical sequences and were confirmed as bovine strain [T. foetus after culture. Nevertheless, DNA sequence of the PCR amplicon showed 99.62% identity with Honigbergiella sp [T. foetus by DNA sequencing, which is especially important in the T. foetus-free or eradicated regions/countries for maintaining such a status.Clinical isolates of e strain . It is wiella sp , which miella sp . TherefoT. foetus, the WSVL had been testing cows with abortion and aborted fetuses for the pathogen. Both cell culture and conventional PCR were employed. In a span of 11 years from 2000 to 2010, 9.7% (9/93) such cows were tested positive, and 4.5% (1/22) of aborted fetuses were similarly tested positive. These cows may play a role in maintaining endemicity of bovine trichomonosis in the corresponding herds [In addition to routine testing bull for ng herds  based ond 4.5% 1/ of abortT. foetus although it started the test in 1970s. Wyoming state rules on bovine trichomonosis (Chapter 15) has been into effect since 2000 with the Wyoming Livestock Board’s enforcing them. State laws mandate bulls grazing on open/public allotments or being traded or leased for the reproductive purpose be tested prior to breeding or change of ownership. They are usually tested in the winter season prior to their being put on pasture the next spring. In 1999, the year before state’s rules of the Chapter 15 came into effect, the statewide prevalence of individual cattle was 2.69%. There was a slow but steady decline since then. In 2008, the statewide prevalence was reduced to 0.62%, a 77% reduction in perveance in nine years. However, the statewide prevalence reached 1.29% in 2009, a 100% increase than the previous year, before it set back to 0.21% in 2010. Based on the data, a linear regression was established as: Prevalence F(x) = 261.020047–0.129685*Year(x)  [T. foetus in bulls if the model were allowed to be used beyond the range of years of its data collection. This had not happened. In next five years from 2011 to 2015, the statewide prevalence was between 0.12 and 0.5% with over 10 000 bulls tested annually. In 2016, the statewide prevalence dropped to 0.03% with only three positive bulls. Further in 2017, 2018 and 2019, three years in a row, not a single T. foetus-positive bull was found throughout the entire state  AI was scarcely used by Wyoming cattle producers. (2) Very low percentage of bulls had been tested three times. The state must continuously enforce its rules since all its adjacent states are still endemic with bovine trichomonosis. In total 33 US states have similar state rules in effect . These rT. foetus-positive bulls is necessarily carried out without any reservation. This is also indirectly supported by the data generated from La Pampa, Argentine. La Pampa is an administrative body in Argentine like Wyoming is a state in the USA. It started a mandatory testing and culling program in 2008. The positive rate of bulls reduced to 1.02% in 2011 from 2.34% in 2008, a 56.4% decrease in only four years. However, the rate remained flat in the next four years between 2012 and 2015. Not culling positive bulls was to be blamed for this flat persistence of bovine trichomonosis [T. foetus-positive bulls besides of AI will lead to total control and the final eradication of bovine trichomonosis. Further, its complement with several other measures such as keeping closed herds, using bulls of three years old or younger and well-fenced allotments will accelerate eradication (Figure Can this success be repeatedly achieved in other US states and rest of the world? To control the disease for minimizing economic losses regulations and rules/laws similar to those of Wyoming are in effect in other 33 US states . To achiomonosis . In addiomonosis . Collect


It has primarily been reported that infusion of herbs as aromatic drinks or tea can protect and heal our organism. From the scientific point of view, it is known that water-soluble substances  have beneficial effects on people’s health. Researchers have taken advantage of plant metabolites as a fundamental part of the healing process, consuming them to treat diseases or relieve pathological conditions. Then, once consumed, these molecules can interact with different targets (including cell membranes), and in some cases, they can even be internalized by cells triggering intracellular responses. The continued search for potential compounds against therapeutic targets is still necessary. Fast screening Informatics’s role in biomedical science has become not a supplementary tool but a necessary support nowadays since it allows researchers to optimize their work in search of new molecules or new properties of well-known molecules. The maturity attained by Quantum Chemistry, Molecular Dynamics, and Molecular Mechanics provides helpful guidance for activities performed by experimentalist scientists, providing support to their scientific investigation that concerns the health arena. The present Research Topic exposes some examples of this theoretical and experimental combined work to reveal relevant properties that secondary metabolites might present.Byrsonima intermedia and Serjania marginata found to possess antiseptic, antimicrobial, anti-haemorrhagic, cicatrizant, and anti-inflammatory properties, computational peptidology applied to determination of the chemical reactivity and bioactivity properties of plant cyclopeptides isolated from Rosaceae, and antioxidant in silico prediction of moracin-C and iso-moracin-C isomers against the OOH free radical. Annona muricata, a plant that has been reported to demonstrate significant antiviral properties against the human immunodeficiency virus, herpes simplex virus, human papilloma virus, hepatitis C virus and dengue virus, to in silico search of GSK3-β inhibitors as a potential Alzheimer’s disease treatment.Overall, this topic covers contributions that range from analytical chemistry to computational chemistry, applied to endemic Brazilian plants such as Thanks to the studies shown in this section, we hope that more researchers will join the search for new properties of biomolecules extracted from different parts of endemic plants. In this sense, Latin America, particularly South America, has enormous potential due to its rich biodeversity that invites scientists to continue the search of these biomolecules with biomedical and biotechnological application.


Clay soils adsorbed the least virus but had the greatest decrease in the river water inoculum (undetectable in ≤ 1 hr), and there was no desorption. Virus desorbed from the other soils into the surrounding water at different rates dependant on soil type . When desorption was no longer detectable, virus persisted, adsorbed to the soil and remained infectious . The long survival of the viruses, particularly at cooler temperatures, highlights the risk of survival in the environment and waterborne spread. The data presented here are highly relevant for assessing risk of pathogen introduction via fomites (stainless steel) and for deciding on best control measures in the context of disease outbreaks.Viral haemorrhagic septicaemia virus (VHSV) and infectious haematopoietic necrosis virus (IHNV) are important viral pathogens posing a serious threat to salmonid fish. Survival of two isolates of IHNV and one of VHSV was assessed at temperatures ranging from 4 to 25°C: (a) after drying on stainless steel, (b) in cell culture medium, (c) in filtered river water, (d) in unfiltered river water, and (e) survival, adsorption and desorption in river sediment and five typical soil types. The viruses survived 1 hr to > 84 days depending on the conditions. Survival was inversely related to temperature and organic and inorganic content. Both viruses remained infectious after being dried on stainless steel for several weeks highlighting the risk of mechanical transmission and persistence in a dry environment. Both adsorbed to the soils from the river water inoculum, with titres between 5.56x10 Both pathogens are notifiable to the World Organization for Animal Health OIE, , and speOncorhynchus mykiss, in Europe and more recently also in a wide range of freshwater species in North America . VHSV has a wide number of hosts; it is currently known to have been isolated from approximately 80 fish species in the Northern Hemisphere and has caused significant losses in farmed rainbow trout, . VHSV haOncorhynchus and Atlantic salmon . Other salmonid species and a range of non‐salmonid species have occasionally been found to be infected in the wild or shown to be somewhat susceptible by experimental infection  was originally identified as a disease mainly of salmon and trout in enzootic areas of western North America, caused by the virus, IHNV. IHNV has since spread and is present in Europe, North America, Russia and several South East Asian countries. Experimental trials have demonstrated IHNV can produce mortality from 3°C to 18°C . With both fish species being highly susceptible to both viruses and with no treatment available, VHSV and IHNV outbreaks could have major economic impact on recreational angling and aquaculture businesses rearing salmon and rainbow trout. The European Reference Laboratory for Finfish Diseases collates data on disease outbreaks in Europe, and conservative estimates indicate that 75 and 46 salmonid farms were classed as ‘known to be infected’ for VHSV and IHNV, respectively, in 2017 (retrieved from http://www.eurl‐fish.eu/Activities/survey_and_diagnosis 16 March 2020).Rainbow trout and salmon are the dominant freshwater salmonid species farmed in Europe and North America. There was approximately 850,000 tonnes of global aquaculture production of rainbow trout worth nearly 4,000 M USD and 2.6 M tonnes of salmon worth 18.5 M USD 1,000 in 2018  after drying on stainless steel, (b) when mixed with cell culture maintenance medium (MM) (positive control), (c) in filtered river water (FRW), (d) in unfiltered river water (URW) and (e) survival, adsorption and desorption in river sediment  and five typical soil types. All mixtures were incubated at a range of temperatures from 4 to 25°C, a range to include both suboptimal and optimal temperatures for survival and infection. Because of the known effects of possible contamination, we also monitored the presence of fungi and bacteria visually during each experiment.Phylogenetic analysis of VHSV has revealed four genogroups, I to IV, which have been divided into further subgroups  cells , originating from common carp Cyprinus carpio, but shown to be contaminated with cells from fathead minnow (FHM) Pimephales promelas  was propagated in ® CCL‐42™), in MM , 1% MEM non‐essential amino acid solution (100x) and 1% penicillin‐streptomycin). .IHNV 32/87 (genogroup E) was propagated in EPC cells in MM , 1% L‐glutamine (200 mM solution), 0.16% sterile tris solution (120 g/L), 0.5% sterile 7.5% sodium bicarbonate and 1% penicillin‐streptomycin). IHNV HV‐90, (genogroup M), was propagated in FHM cells , and the supernatant containing the cultured virus was stored at 4°C and used within two weeks of harvest.Virus stocks were prepared by propagating the viruses (previously stored at −70°C) and harvesting from monolayers showing full cytopathic effect (CPE). The harvests were clarified by low speed centrifugation (2000 x 2.250/ml) were calculated , and serial 10‐fold dilutions were made down the plate. Titrations were incubated in a humid chamber for 7 days at 15°C and end point 50% tissue culture infectious doses , IHNV 32/87 (with an average titre [n = 2] of 1.48 × 108 TCID50/ml) and IHNV HV‐90 (with an average titre [n = 2] of 3.38 × 106 TCID50/ml) was added to autoclaved stainless steel discs, 1 cm in diameter  in October (when IHNV testing), September (when VHSV testing) and February (when testing both viruses for adsorption and desorption) . RW samples were unfiltered and filtered through a 0.2 µm filter with cellulose acetate membrane. Sediment was collected from the side of the riverbank where there was wet mud. Large stones and plant materials were removed; the sediment was sieved (1.4 mm mesh), and the sample centrifuged at 1,500 × 8 TCID50/ml for VHSV J167; 5.56 × 107 TCID50/ml for IHNV 32/87 and 1.20 × 108 TCID50/ml for IHNV HV‐90. A 1 in 10 dilution of virus was created by adding 5 ml stock virus to the MM and FRW, and 10 ml of the stock virus was added to the URW to create a 1 in 5 dilution. A 50 g sample of river sediment was mixed with 50 ml URW containing stock virus at a 1 in 5 dilution. Preparations were returned to their respective temperatures for incubation and sampling.A single closed glass bottle containing either 45 ml of MM, 45 ml of FRW, 40 ml of URW or 50 g of river sediment was equilibrated overnight at each of five temperatures: 4, 10, 15, 20 and 25°C. The following day, virus suspensions were made in each matrix from stock virus stored at 4°C. Starting titres of stock viruses were determined as 3.78 × 10g for 15 minutes at 4°C and the supernatant filtered (0.2 µm cellulose acetate membrane) before titration. Mixtures were replaced back in their allocated incubators immediately after sampling. Once the titre of virus in a test matrix was no longer detectable for two consecutive weeks, titrations ceased for that matrix‐temperature combination. The limit of detection of the assay was 1.76 × 10−1 TCID50/ml.Samples were taken at day 0 (1 hr after spiking), days 1, 2, 3 and 7, then weekly intervals for up to 72, 65 and 84 days for IHNV 32/87, IHNV HV‐90 and VHSV J167, respectively, to assess virus survival over time. On two occasions (for IHNV 32/87 after day 56 and IHNV HV‐90 after day 49), the weekly sample interval was extended to 9 days. At each sampling, matrix suspensions were vortexed. For spiked MM and FRW, 20 µl aliquots were titrated directly onto cells as described in virus quantification above. 0.5 ml aliquots from the spiked URW were filtered (0.2 µm cellulose acetate membrane) immediately prior to titration to prevent any interfering microbial growth on the cells. Spiked sediment was mixed by manual agitation (inversion and swirling), 2 ml aliquots were centrifuged at 2060 × 2.58 TCID50/ml), (b) IHNV 32/87 (with starting titre 1.76 × 107 TCID50/ml) and (c) IHNV HV‐90 (with starting titre 2.58 × 107 TCID50/ml) and 16 ml FRW (equilibrated to 4°C). The mixtures were incubated at 4°C, and after 1 hr, 3 hr, 1 day, 7 days and then weekly, the mixtures were vortexed and the combined sediment‐FRW and soil‐FRW mixtures titrated directly to quantify the virus survival. The mixture was then centrifuged  and the supernatant filtered (0.2 µm cellulose acetate membrane) and titrated. The supernatant was replaced with FRW , after each titration to observe any desorption of virus from the soil into each wash. The same FRW was run alongside the experiment, 4 ml each virus was added to 16 ml FRW (1 in 5 of each virus) and maintained under the same conditions with the same sample points.Five dry typical soil types, clay soil, clay loam, chalk soil, chalk loam and sandy soil (Fisher Scientific™ UK), were autoclaved and ground in a pestle and mortar to remove large lumps and allow homogenous particle mixing Figure . The riv‐2 TCID50/ml for the removed liquid but visibility was restricted for the soil mixture meaning the limit of detection was between 1.76 × 10−3 TCID50/ml and 2.58 × 10−5 TCID50/ml.Once the titre of virus in a test matrix was no longer detectable for at least two consecutive weeks, titrations ceased. The limit of detection of the assay was 1.76x1010 converted to calculate the log10 drop from that adsorbed in the first hour.Initial adsorption, within the first hour, was calculated by removing the titre of the virus in the FRW inoculum from the titre in the FRW‐soil/sediment mixture at 1 hr. Survival of the adsorbed virus in the soils/sediment was also measured by ‘the titre of the virus in the combined FRW‐soil/sediment mixture’ minus ‘the titre of the desorbed virus’. This was log3The survival of VHSV and IHNV over time in cell culture medium, filtered and unfiltered river water, associated with sediment and dried on stainless steel at various incubation temperatures is represented in Figure 3.110 drop) after the maximum time tested (65 to 84 days). The shortest survival in all matrices, except after drying on stainless steel, was at 25°C; viruses were undetectable at the end of the study . The maximum survival at 25°C was in MM or FRW: 49 to 56 days, 35 to 42 days and 42 to 49 days for VHSV J167, IHNV 32/87 and IHNV HV‐90, respectively.The survival times of both VHSV and IHNV decreased as temperature increased in all matrices, excluding virus dried at 15°C on stainless steel Figure . The lon3.210 drop) between 42 and 49 days and this was also the case at 15°C. IHNV 32/87 decreased the fasted at 15°C and was no longer detected at day 35 (5.92 log10 drop) but at 10, 20 and 25°C there was a similar rate of decrease and virus was still detectable at day 56. IHNV HV‐90 had a similar rate of decrease at temperatures between 10 and 25°C; virus was undetectable at day 7 and 14 (4.28 log10 drop).Survival of all virus isolates was longer when dried on the stainless steel than in the URW and the URW‐sediment mixture Figure . The sur3.3Survival at 4 to 25°C in MM ranged between 49 and > 84 days for VHSV J167, 21 to > 72 days for IHNV 32/87 and 42 to > 65 days for IHNV HV‐90 Figure . Surviva10 of the calculated starting titres 1 hr after spiking, whereas a drop of between 2.47 and 5.47 log10 was seen in the water that was separated from the unsterile URW‐sediment mixture. All viruses were undetectable after 1 day in water that was separated from the URW‐sediment at 25°C for all viruses . At 4 and 10°C, the survival only dropped below detectible levels between 42 and 49 and 14 to 21 days, respectively, for VHSV J167. The decrease was more rapid for IHNV: 3 to 7 and 1 to 2 days, respectively, for IHNV 32/87 and 14 to 21 and 7 to 14 days, respectively, for IHNV HV‐90.Survival was lower in URW compared to FRW at all temperatures tested. However, titres of virus in FRW and URW samples were within 1 logFungal growth was first visible at the higher temperatures: in URW fungus was present from day 2 at 20 and 25°C, day 3 at 15 and 10°C and day 14 at 4°C. FRW had some fungal growth visible at day 21 at 25 and 20°C and day 49 at 15°C but no fungus at 10 and 4°C. Fungal growth was observed from day 28 at 15°C in the MM inoculated with IHNV 32/87. The results show the viral titre in the MM drop below detectible levels quicker than FRW in this instance Figure . No fung3.4Figure . Figure 3.510), was within 1 hr for clay soil and clay loam. After calculating the quantity of adsorbed virus, it was found that virus that adsorbed to clay soil was still infectious but had a 1.33 to 2.5 log10 drop in titre after 1 hr compared to that in the spiked FRW wash  at time 0. After replacing the FRW, there was no desorption of virus into the FRW washes. The FRW‐soil/sediment mixtures of non‐clay‐based soils had a higher titre of adsorbed virus after 1 hr (≤ 1.17 log10 drop from the inoculum) even though less virus appeared adsorbed than the clay soils; 8.15x102  to 3.78 × 106  remained unadsorbed in the FRW inoculum separated after 1 hr.After 1 hr, there was an immediate drop in viral titres in the FRW inoculum separated from the five sterile soil and sediment mixtures compared to higher levels in the unseparated soil‐liquid mixture Figure . The hig3.610 drop visible was between 2.67 and 4.33 between the viruses from a possible 3.5 to 7.17 log10 drop from the absorbed virus. Based on the visible rate of decrease it is likely viable virus remains in all soil types, beyond the point at which it was no longer detectable. Although starting with a higher titre adsorbed to the soil, there were steeper drops in the non‐clay soils and sediment. Based on the rate of decay, it is likely the virus in clay soils will survive the longest, but the detectable limit would have to be lower to confirm this.Titres slowly decreased in the soil for all isolates, and the VHSV J167 isolate was detected longer than the IHNV isolates in all soils Figure . The lon3.7Apart from VHSV and IHNV adsorbed to clay soil and IHNV adsorbed to clay loam, the other soils and the sediment showed desorption into the weekly washes of FRW (but with decreasing amounts each week) Figure . The lon4As environmental conditions vary considerably and there are discrepancies between published data, continued research on the persistence of viruses is necessary for informing disease control risk assessments and emphasise the relevance of strict biosecurity protocols to prevent introductions. Our results assist in one of the evaluating requirements for virus transmission via water published by Oidtmann and colleagues , statistical significances between our findings cannot be provided. This limits the comparisons that can be made between treatments and each virus; however, descriptive data are provided, and qualitative comparisons can be made.VHSV and IHNV can be very sensitive to ultraviolet light, even in aqueous solutions with high turbidity  at temperatures between 37°C and 4°C) Ahne, , and surIn this study, survival of both VHSV and IHNV after drying on stainless steel was longer than under the natural environmental conditions mimicked by URW and the URW‐sediment mixtures. The inverse relationship between temperature and survival observed in the other matrices was apparent for VHSV, except for the 15°C samples that decreased at the same rate as the 25°C samples. The IHNV isolates showed very similar survival rates between 10 and 25°C Figure . Without4.3Cell culture MM was used as a positive control representing most favourable conditions. It provides an indication of the stability of the viruses but unlikely to provide realistic data when relating to survival in the environment. Survival in freshwater was investigated for its relevance to the UK and because it is the most common environment of rainbow trout and the majority of other species susceptible to VHSV and IHNV. Survival in FRW was included to give a ‘worst‐case’ scenario in water, that is how long the virus may survive in clear water under ideal conditions, compared to a more relevant survival time in URW containing organic and inorganic content and live biota.10 drop for all viruses in 35 to ≥ 65 days at 15°C compared to 1 log10 drop within 24 hr at 14°C   at temperatures ranging between 4 and 25°C. Viral titres decreased quickest at all temperatures in the water sample separated from the URW‐sediment. The sharp drops in virus titres in the separated liquid, 0 to 1 hr after addition of the virus to sediment at all temperatures tested Figure , suggestThere are no studies to compare VHSV inactivation in different sediments. One study has been published providing data on the survival of VHSV in a cell culture medium‐unsterile pond sludge mixture by quantifying the virus in the separated medium Ahne, , and twohttps://www.iscc‐system.org/wp‐content/uploads/2018/09/Clay‐sandy‐and‐peat‐soils‐and‐soil‐organic‐matter.pdf accessed 28 August 2020). Incubation was at 4°C to give a worst‐case scenario within the range of temperatures already studied. An elution process such as that published for enteric human viruses . The ‘adsorbed’ virus presented in these soils is therefore decreasing due to a combination of desorption and inactivation. The differing quantities desorbed imply the soil type is a determining factor in viral desorption. It should also be remembered that some desorbed virus may also have been inactivated during the weekly sample points.The non‐clay‐based samples had higher levels of virus left in the FRW inoculum after 1 hr of mixing, but higher levels of adsorbed virus calculated in the soils and sediment than the clay‐based soils, so the initial inactivation seen in the clay‐based soils is not evident. The soils adsorbed virus with ≤ 1.17 loghttps://www.iscc‐system.org/wp‐content/uploads/2018/09/Clay‐sandy‐and‐peat‐soils‐and‐soil‐organic‐matter.pdf accessed 27 August 2020). Chattopadhyay and Puls suggested that viruses adsorb to colloidal clays using van der Waals forces. The pH influences the charge on both viruses and soils, and ionic strength and its constituents determine the binding forces between them  and because of this, initial adsorption may have been less as the water stays on top of the soil and did not move inside as easily. Desorption is less likely because the water moves through very slowly, so the clay soils have a very good water holding capacity . Sandy soils have relatively large particles (0.05 to 0.2 mm) so could explain the opposite effect, high starting adsorption with a long desorption and rapid decrease in adsorbed virus. Loamy soils have a mixture of particles so would sit somewhere in the middle—this is less clear with the results but may have become more apparent over time (10 decrease of adsorbed virus), virus in the clay soils is likely to have survived the longest, and there appears to be differences in the decay depending on soil type that may have been more or less pronounced over time. Survival and adsorption may also differ between viruses; this also may have been more evident over time. Previous studies on the adsorption of different viruses to soils have shown a high degree of variability not only among different virus types but among different strains of the same virus type  prevents us knowing how long survival was past the last detection. Our detection of all the adsorbed viruses is likely to be an underestimate of survival based on the rate of decay; however, the rate of decay is useful for disease models. Based on the rate of decay  may appear to be an appropriate method for waste water sanitation from processing facilities, as concluded by Skall and co‐workers for VHSV ; in the water separated from URW‐Sediment mixtures VHSV survived 1 hr to 49 days and IHNV 1 hr to 21 days, when dried on stainless steel VHSV survived 42 to > 49 days and IHNV 3 to > 56 days, at temperatures ranging from 25 to 4°C. Within the tested range, the lower half of the temperature range has the longest survival and is particularly relevant when considering water temperature in relation to UK climate and deeper water. This study has demonstrated that a range of environmental parameters influence the survival of viruses and machinery and surfaces made of stainless steel should be cleaned and disinfected to prevent potential spread of disease. Persistence of VHSV J167, IHNV 32/87 and IHNV HV‐90 was strongly dependent on temperature, presence of organic and inorganic content, live biota and type of soil association, and although not tested here, it is known that UV is a strong inactivating factor even in turbid water  virus strain; (b) viral shedding rates per animal or biomass per time unit; (c) prevalence of infection in the shedding aquatic animal population over the course of a disease outbreak; (d) water flow rates, which will dilute virus concentrations; and (e) minimum infectious dose for exposed naive fish (Oidtmann et al., The authors confirm that they have no conflict of interest to declare.The authors confirm that the ethical policies of the journal, as noted on the journal's author guidelines page, have been adhered to. No ethical approval was required as this study was in vitro and no animals were used in this study.Table S1Click here for additional data file.Table S2Click here for additional data file.


Complications lead to unplanned readmissions (UR) and are reported to be associated with a two- to threefold increase in hospital admission costs. Since healthcare costs are increasing worldwide, cost containment is the major challenge for future healthcare. In the literature, there are only a few studies that analysed hospital costs after pancreatoduodenectomy (PD). In this study, we aimed to create an understanding of the costs of complications and UR in patients who underwent a PD.p = 0.001). The median costs for minor complications were EUR 30.316, and for major complications, EUR 42.664 (p = 0.001). Costs were related to the Comprehensive Complication Index (CCI). The median costs of patients with one or more UR were EUR 41.199. Conclusions: Complications after PD led to a EUR 4.634–EUR 16.982 (18–66%) increase in hospital costs. A UR led to a cost increase of EUR 12.567 (44%). Since hospital costs are directly related to the CCI, reduction in complications will lead to cost-effectiveness.Background/Objectives: Complications after pancreatoduodenectomy (PD) lead to unplanned readmissions (UR), with a two- to threefold increase in admission costs. In this study, we aimed to create an understanding of the costs of complications and UR in this patient group. Furthermore, we aimed to generate a detailed cost overview that can be used to build a theoretical model to calculate the cost efficacy for prehabilitation. Methods: A retrospective cohort analysis was performed using the Dutch Pancreatic Cancer Audit (DPCA) database of patients who underwent a PD at our institute between 2013 and 2017. The total costs of the index hospital admission and UR related to the PD were collected. Results: Of the 160 patients; 35 patients (22%) had an uncomplicated course; 87 patients (54%) had minor complications, and 38 patients (24%) had severe complications. Median costs for an uncomplicated course were EUR 25.682, and for a complicated course, EUR 32.958 ( A pancreatoduodenectomy (PD) is a complex procedure usually performed in cases of malignant tumours of the pancreatic head or surrounding structures such as the duodenum, the ampulla of Vater, or the distal bile ducts. Over the past decades, the mortality of pancreatic resections has been reduced to less than 5% by centralization of care. However, the overall morbidity of pancreatic resections is still high, ranging from 30% to 60% ,2,3. TheWith increasing age and multimorbidity, patients are also prone to nonsurgical complications such as pneumonia and cardiovascular events. Complications and the often necessary readmissions are both associated with high hospital costs. In the literature, the median costs for patients undergoing a PD without complications are between EUR 17.000 and EUR 30.000 ,7,8,9 FoThe Comprehensive Complication Index (CCI) seems directly related to hospital costs . The CCISince healthcare costs are ever-increasing worldwide, one of the major challenges for the future is balancing costs and potential survival gain, taking quality-adjusted life years into consideration. The primary aim of this study was to gain understanding of the impact of complications and unplanned readmissions, considering the CCI score, on perioperative hospital costs in patients who underwent a PD. Furthermore, we aimed to generate a detailed cost overview that can be used to build a theoretical model to calculate the cost efficacy for a multimodal prehabilitation program .Since July 2013, all patients in the Netherlands who underwent a pancreatic resection have been registered prospectively in the Dutch Pancreatic Cancer Audit (DPCA). This study is a retrospective cohort analysis using the DPCA database of patients who underwent a pancreatoduodenectomy at our tertiary referral center, the University Medical Centre Groningen (UMCG), between 2013 and 2017. In total, 160 patients were included. Pre-, intra- and postoperative variables were obtained. All consecutive adult patients who underwent PD (classic Whipple procedure/pylorus-resecting pancreatoduodenectomy (PRPD) or pylorus-preserving pancreatoduodenectomy (PPPD)) between 2013 and 2017 at our institute were included in the study.www.assessurgery.com (accessed on 3 October 2021) [All treatment-related incurred healthcare costs of the index hospital admission and unplanned readmission (UR) related to the PD, from one day before PD until the date of discharge, were obtained from the financial department. These healthcare costs included all components of the surgical procedure, postoperative in-hospital care, postoperative hospital visits within 90 days after discharge, and in-hospital rehabilitation program, and concerned the actual individual patient-related costs that our institute incurred to treat the specific patient for PD. Therefore, a prolonged stay at the ICU or a longer surgical procedure resulted directly in higher costs. The costs for preoperative work-up were not included. For further analysis, the total costs were subdivided into seven cost domains: general diagnostics , imaging , outpatient clinic , clinical care (ward care), surgical , ICU , other costs . Analysis was carried out using 2017 costs in euros (EUR). Complicated postoperative course and complications were graded by the Clavien–Dindo classification ,13. The er 2021) . The prit-test was used. A Kruskal–Wallis test was used if data was not normally distributed to analyse continuous variables for multiple groups. In case of nominal/categorical variables, a Chi-squared test or a Fisher’s exact test was used; p-values < 0.05 were considered significant. Analyses were performed in SPSS v.24 Data were collected in Microsoft Office Excel 2010 and later transferred to SPSS Statistics 24  for statistical analysis. Cost analysis was performed using R 3.6.1 . Descriptive statistics were calculated as percentages, median, or mean, as appropriate. For the cost calculations, the interquartile range was used. A Mann–Whitney test was used for univariate analysis of continuous variables if data were not normally distributed. In case the data were normally distributed, an unpaired All data were retrieved from an existing database from the Dutch Pancreatic Cancer Audit (DPCA). Since we used the existing data register from the DPCA from the patients of our own hospital, no formal approval from the medical ethics committee was needed. Our local ethics committee waived the need for informed consent.A total of 160 patients  who underwent a PD between January 2012 and November 2017 were included for analysis. A total of 125 (78.1%) of the patients underwent a PPPD, and 35 (21.9%) patients underwent a classic Whipple procedure. A vascular resection was performed in 26 patients (16.3%). In total, 97 patients (62.9%) underwent preoperative biliary stenting. In 80 patients (50%), the tumor originated from the pancreas. In 120 patients (70%), the pathology showed an adenocarcinoma. In total, 12 patients (7.5%) developed a clinically relevant (grade B/C) postoperative pancreatic fistula (CR-POPF) (see p = 0.001). For patients with minor complications, the median costs were EUR 30.316 (IQR EUR 24.717–EUR 36.291). For patients with major complications, these costs were EUR 42.664 . For patients with Clavien–Dindo grade IV complications, the median costs were EUR 90.0841 (IQR EUR 49.646–EUR 116.845). Complications after PD led to an increase in total hospital admission costs of EUR 4.634 (18%) in the case of minor complications; and in the case of major complications, EUR 16.982 (66.1%) (p = 0.001). (The median costs for patients with an uncomplicated course after PD were EUR 25.682 (IQR EUR 21.447–EUR 31.594). For patients with a complicated course  after PD, the median costs were EUR 32.958 . .p = 0.001). Surgical costs seemed to be more or less the same for patients suffering no complications, minor complications, or major complications, and ranged from EUR 10.930 to EUR 12.210 (p = 0.642)  .p = 0.001–0.023). ICU costs, general diagnostic costs, imaging costs, and other costs seemed to relatively rise the most in case of Clavien–Dindo grade IV complications. . The mean and median days of hospital stay for patients with minor complications were 17.1 days and 15 days, respectively. For patients with major complications, the mean and median days of hospital days were 37.6 days and 31 days, respectively (p = 0.001)  after PD, the mean and median days of hospital days were 23.3 days and 17 days, respectively (= 0.001) .p = 0.001). The median costs for every different category rose when the CCI score increased. With clinical care costs, ICU costs and imaging costs increased the most in cases in which complications added up (n = 6)  . Figure p = 0.001)  .p = 0.001). For patients with UR and minor complications, the median costs were EUR 40.875 (IQR EUR 30.473–EUR 43.260). For patients with UR and major complications, these costs were EUR 41.739 . A UR was associated with an increase of EUR 12.567 (44%) in total hospital costs compared to the group of patients without readmissions (p = 0.001). This increase was mainly explained by an increase in general diagnostics costs, imaging costs, clinical care costs, and other costs, as these costs were higher in cases of a UR (p = 0.001–0.002)  . The medp = 0.001)  .This single academic center study revealed that complications after PD led to a EUR 4.634–EUR 16.982 (18–66%) increase in hospital admission costs. These costs were mainly due to an increase in clinical care costs. In cases of Clavien–Dindo grade IV complications, the median costs in every cost domain were higher compared to an uncomplicated course or less-severe complications (Clavien–Dindo I–III). In particular, ICU costs, general diagnostic costs, imaging costs, and other costs seemed to relatively rise the most compared to less-severe complications (Clavien–Dindo I–III). A UR was associated with a EUR 12.567 (44%) increase in hospital costs. Log hospital costs seemed to be directly related to the CCI score, with a Pearson correlation coefficient of 0.77. Clinical care costs, ICU costs, and imaging costs especially seemed to rise the most in case complications add up.With healthcare costs increasing annually worldwide, hospital cost containment will be one of the biggest challenges. One of the reasons for the increase in healthcare costs is the ageing population, resulting in patients with multimorbidity, and more often a complicated course after surgery. Postoperative complications have a high impact on quality of life and healthcare costs for society. A PD is a complex procedure often performed in specialized pancreatic centres. Although the mortality rate of pancreatic resections has been reduced over the past decades to less than 5% by means of centralization, leading to higher volumes and lower failure to rescue rates, the overall morbidity remains around 30% to 60% ,8 Well-kThe following three strategies could reduce costs. Early detection and treatment of complications might prevent worsening of severity of complications. In 2018, the DPCG started the PORSCH trial, a nationwide stepped-wedge cluster randomized trial . The aimAnother strategy to reduce complication and thereby lower healthcare costs might be the introduction of surgical care bundles (SCBs) and enhanced recovery programs (ERPs) in pancreatic surgery. Over the past decade, the introduction of SCBs in colorectal and liver surgery have proven to significantly reduce surgical site infections (SSI) ,17. VariA third strategy to reduce complication and prevent worsening of severity of complications and thereby lower healthcare costs might be prehabilitation. Prehabilitation is a way to preoperatively optimize modifiable risk factors such as poor physical fitness, malnutrition, low mental resilience, iron deficiency anaemia, and smoking and/or drinking alcohol ,24,25. TOur study had potential limitations. Only healthcare costs incurred in our institute were available for analysis. Since our hospital is a tertiary referral center for pancreatic surgery, some patients might have been readmitted in a local hospital nearby instead of the index hospital. This means that some costs might not be included in our study, meaning the overall costs of a complications and UR after PD might be even higher than previously stated. Nevertheless, we expect that the number of patients who did not go to our hospital for their complication was low, since patients were advised to go to the index hospital in case problems occurred. Another limitation was that we only examined hospital costs. Costs from an eventual admission in a nursing home or rehabilitation center after hospital discharge were not included. This means that the overall costs of a complications and UR after PD might be even higher than previously stated.The strength of our study was that we used a larger study population (160 patients) and a more detailed hospital costs analysis compared to previous studies. Furthermore, our study and detailed cost overview served as a substantiation for a business model for a multimodal prehabilitation program in our hospital .Our study showed that complications after PD led to a EUR 4.634–EUR 16.982 (18–66%) increase in hospital admission costs, mainly due to an increase in costs of clinical care. A UR was associated with a EUR 12.567 (44%) increase in hospital costs. Hospital costs seemed to be directly related to the CCI score. Therefore, every reduction in postoperative complications will contribute to an increased cost-effectiveness.


Visacane is a sugarcane quarantine station located in the South of France, far away from sugarcane growing areas. Visacane imports up to 100 sugarcane varieties per year, using safe control and confinement measures of plants and their wastes to prevent any risk of pathogen spread outside of the facilities. Viruses hosted by the imported material are either known or unknown to cause disease in cultivated sugarcane. Poaceae viruses occurring in plants surrounding the quarantine glasshouse are currently unknown. These viruses could be considered as a source of new sugarcane infections and potentially cause new sugarcane diseases in cases of confinement barrier failure. The aim of this study was to compare the plant virome inside and outside of the quarantine station to identify potential confinement failures and risks of cross infections. Leaves from quarantined sugarcane varieties and from wild Poaceae growing near the quarantine were collected and processed by a metagenomics approach based on virion-associated nucleic acids extraction and library preparation for Illumina sequencing. While viruses belonging to the same virus genus or family were identified in the sugarcane quarantine and its surroundings, no virus species was detected in both environments. Based on the data obtained in this study, no virus movement between quarantined sugarcane and nearby grassland has occurred so far, and the confinement procedures of Visacane appear to be properly implemented. Saccharum interspecific hybrids) belongs to the Poaceae family and is cultivated in tropical and subtropical areas for the production of sugar and ethanol. To be competitive on the global market and for diversification of products, sugarcane farmers need to use the best-performing varieties and breeders need genetic resources to create them. Consequently, moving plants across international borders for exchange between breeders or directly to producers is an essential step of this process. Plant movement involves biosafety to protect local production by avoiding pathogen introduction and spread. Safe and accurate quarantine procedures need to be implemented, including accurate disease testing [Sugarcane . Nowadays, Visacane works with research organizations and sugarcane companies from tropical and subtropical locations around the world [To control disease introduction through sugarcane germplasm movement in the outermost French European regions and for partners in Africa, a sugarcane quarantine station was established in 1971 by the Institut de Recherche en Agronomie Tropicale in Nogent sur Marne, France . This quhe world . Importehe world ,5,6 thathe world ,6,7,8,9.Mastrevirus (Geminiviridae family), Tritimovirus (Potyviridae family) and Luteovirus (Luteoviridae family) [Polerovirus (Luteoviridae), Potyvirus (Potyviridae) and Mastrevirus (Geminiviridae) cause diseases in sugarcane [Because Visacane is located outside a sugarcane growing area, the risk of sugarcane infection by surrounding local viruses should be nonexistent or minimal. Nevertheless, the purpose of quarantining is also to avoid the release or escape from quarantine of viruses that could potentially threaten the local vegetation. Infrastructures and processes are rarely 100% safe and a flow may exist between a quarantine glasshouse and its surrounding environment. Importation of infected plants in a new area is a risk for disease emergence and newly introduced viruses may invade communities of native plants . Maize y family) ,19,20. Augarcane .Consequently, to investigate the potential flow of plant viruses between a quarantine and its environment, deciphering the plant virome of both entities is necessary. At the present time, the best methodology for virome identification in a specific environment is viral metagenomics, and different approaches are available ,22. ViriThe youngest fully developed leaf  was sampled from 20 sugarcane plants representing 19 diseased varieties currently maintained in the quarantine glasshouse of Visacane at Cirad in Montpellier . Five grhttps://www.emplantbase.org/home.html, accessed on 31 March 2021). Five grams of leaf and stem tissue from each wild Poaceae were cut in small pieces and stored as described above for sugarcane.Wild Poaceae were collected from three areas surrounding the quarantine glasshouse, including a flat stone land adjacent to the quarantine glasshouse, a sloped land slightly further from the quarantine glasshouse, and a grassland at least 25 m distant from the glasshouse . One hung for 2.5 h at 4 °C to concentrate viral particles. Nonencapsidated nucleic acids were eliminated by DNase and RNase incubation at 37 °C for 1.5 h. Total RNA and DNA was then extracted using the NucleoSpin kit (Macherey Nagel). Reverse transcription was performed with the SuperScript III reverse transcriptase (Invitrogen), cDNAs were purified with the QIAquick PCR Purification Kit (Qiagen) and complementary strands were synthesized using the Klenow DNA polymerase I. Double-stranded DNA was amplified by random PCR amplification. Samples were barcoded during reverse transcription and PCR steps were performed using homemade 26-nt Dodeca Linkers and PCR multiplex identifier primers. PCR products were purified using NucleoSpin gel and PCR clean-up (Macherey Nagel). Wild Poaceae and sugarcane samples were sequenced in two independent Illumina HiSeq runs in order to avoid index-hopping contamination [Each of the 150 collected samples was processed using the virion-associated nucleic acid (VANA)-based viral metagenomics approach . Brieflymination . Bioinfomination . Brieflymination . Adaptormination . The clemination . Putativmination  from sugRT-PCR reactions were carried out to extend the genomic sequence of a novel umbravirus isolated from sugarcane variety BJ790038, for which four short contigs (131–325 nt) were recovered using the VANA-based metagenomics approach. PCR reactions were performed using the Qiagen OneStep RT-PCR Kit. Ten primers were designed  based onhttps://blast.ncbi.nlm.nih.gov/Blast.cgi, accessed on 31 March 2021). Forty-eight contigs assigned to plant viruses obtained from both compartments (within and outside the quarantine) were selected for further comparisons. These 48 sequences were subsequently aligned to reference genomes using ClustalW with default settings [Caulimoviridae, Closteroviridae, Geminiviridae, Luteoviridae, Potyviridae, and Tombusviridae families, that were used for the phylogenetic analyses described above are listed in the Contigs produced by assembly of Illumina reads were utilized as queries to perform BLASTn and BLASTx searches  using thsettings . Visual settings . The JukTombusviridae) to 833,570 (Geminiviridae)  to 625,654 (KN8924), with an average number of 315,508 reads per sample . Retaineviridae) .Helictochloa bromoides from grassland) to 414,878 (Bromopsis erecta from flat stone land), with an average number of 175,553 reads per sample. Retained virus reads represented 2.8% of the 22,821,881 reads obtained for the three sampled locations , one unclassified genus (Sobemovirus) and one unclassified plant-associated virus, Trifolium-associated circular DNA virus , Mastrevirus (Geminiviridae) and Umbravirus (Tombusviridae) genera were present in both quarantined sugarcane samples and wild Poaceae samples.Contigs from quarantined sugarcane varieties were assigned to seven virus families, i.e., sviridae . With thNA virus . ContigsClosteroviridae contigs obtained from seven quarantined sugarcane varieties  were assigned to the Ampelovirus genus, one Closteroviridae contig obtained from a grassland sample of Brachypodium phoenicoides was assigned to the Closterovirus genus using BLASTx searches , suggesting that it represented a novel closterovirus that needs further characterization. The phylogenetic analyses also showed that the sugarcane Ampelovirus contigs grouped together but were also distributed into several subgroups, thus indicating that sugarcane-associated ampeloviruses are diverse and may represent several divergent variants of the same species, or even several species  were assigned to the Luteoviridae family using BLAST searches  ranged from 39 to 43% and from 70.8 to 74.9% with unclassified luteoviruses isolated from insects . The phylogenetic analysis allowed us to confirm that the Luteoviridae contigs obtained from the two wild grasses were highly divergent from plant luteoviruses , which is below the species demarcation (<76% nucleotide identity) of the Potyviridae family [Tritimovirus genus.Complete genome sequences of viral isolates belonging to the imovirus . These rimovirus A, and the family . ConsequAnisantha madritensis plant samples collected from the flat stone land , a mastrevirus recently reported from the Réunion Island. They also shared 44.3–46.2% identity with the SSEV and SWSV genome sequences retrieved from the quarantined sugarcane plant samples. In a phylogenetic tree constructed with entire genome sequences of known mastreviruses and those obtained in this study, the two sequences from the wild Poaceae plants were also located at a unique branch  were obtained from nine sugarcane samples , from which two contigs, each covering the full badnavirus genome, were produced for sugarcane variety BJ790038 and variety X . These wild Poaceae samples included Trachynia distachya, Bothriochloa barbinodis, B. phoenicoides, Phalaris minor, and Festuca sp. plants from both the sloped land and the grassland areas (Umbravirus contigs recovered from wild Poaceae plants shared 45–72% nucleotide identities with a sugarcane umbra-like virus genome (accession number MN868593). Four contigs obtained from two P. minor plants, one Festuca sp. plant, and one B. phoenicoides plant had 66–69% nucleotide identity with strawberry-associated virus A (accession number MK211274). The four contigs also clustered in a neighbor-joining phylogenetic tree, thus suggesting that a single novel umbravirus infected these wild Poaceae plants (Festuca sp. plant shared 71% nucleotide identity with Patrinia mild mottle virus (accession number MH922775), contigs from T. distachya and B. barbinodis were 100% identical and had 71–73% nucleotide identity with Ethiopia-maize-associated virus (accession number MF415880). Finally, two partial genome sequences of umbraviruses , and one unclassified plant-virus (trifolium-associated circular DNA virus). In contrast, only seven families formed the plant virome of the quarantined sugarcane varieties. Most virus sequence reads (85%) obtained from wild Poaceae were attributed to non-plant viruses . Fungal virus reads related to Chrysoviridae, Partitiviridae, and Totiviridae were previously identified in wild plants, including Poaceae [Caulimoviridae, Closteroviridae, Geminiviridae, Luteoviridae, Potyviridae, and Tombusviridae. The sequences obtained herein for the Closteroviridae, Luteoviridae, and Potyviridae families belonged to different virus genera when comparing virus isolates from quarantined plants and from plants growing in the outside quarantine environment. Nevertheless, even if the virus genera between quarantined and non-quarantined plants are different, the risk of cross infection cannot be eliminated. For the contigs assigned to the three other families, the genera were common between quarantined sugarcane and wild Poaceae. These common genera were identified as Badnavirus (Caulimoviridae family), Mastrevirus (Geminiviridae family), and Umbravirus (Tombusviridae family).The plant virome of the wild Poaceae collected near the quarantine glasshouse included 11 virus families, one unclassified genus  and Pellissanne (12 reads), two locations that are, respectively, 30 and 120 km distant from the sugarcane quarantine in Montpellier . However, further investigations are needed to identify the potential risk for sugarcane to be infected by the putative B. phoenicoides badnavirus discovered in this study.Major plant species infected by badnaviruses have a tropical or sub-tropical origin  . Badnavibgroup D , thus inA. madritensis in our study is the first report of this virus. It could be the cause of new diseases of cultivated Poaceae in Europe. This virus is different from currently known mastreviruses of sugarcane and, although genetically distant from sugarcane streak Egypt virus (SSEV) and sugarcane white streak virus (SWSV), it might be able to cause disease in sugarcane in cases of accidental spread to healthy sugarcane plants.Mastreviruses occur in cultivated and wild Poaceae in several continents ,45,46,47B. barbinodis and T. distachya). Specifically, Ethiopia-maize-associated virus was detected in the quarantine glasshouse in the sugarcane variety KN8924 that originated from Sudan. Consequently, umbra-like virus diversity does not appear related to plant species and this type of virus could spread among different plant hosts, such as between plants in quarantine and in surrounding area compartments. Several other umbravirus isolates found in wild Poaceae in this study differed genetically from isolates close to the sugarcane and maize umbra-like viruses. Isolates of these putative novel umbraviruses were distributed in two phylogenetic subgroups suggesting that two unclassified umbraviruses, not related to any currently known sugarcane umbravirus, were circulating among several wild Poaceae species growing near the sugarcane quarantine greenhouse in the South of France. These two viruses may represent two novel species of the genus Umbravirus while the International Committee on Taxonomy of Viruses recommends a species demarcation threshold of <70% nt sequence identity for umbraviruses [Umbraviruses and unclassified umbra-like viruses were the final viruses shared by quarantined sugarcane and wild Poaceae. Umbra-like viruses, which are similar to umbraviruses but do not contain all of the umbravirus genomic features , have beaviruses .The area surrounding the sugarcane quarantine of Cirad in the South of France hosts a great diversity of viruses in wild Poaceae, which appears greater than the one occurring in quarantined sugarcane varieties. As these two environments share viruses from the same family and even from the same genus, the risk of cross contamination seems higher than expected. However, we showed in this study that no identical or highly similar virus sequences were present in quarantined material and in plants growing in its surrounding environment. This suggested that plant virus movement between quarantined sugarcane and plants of nearby locations has not, so far, occurred. While a large majority of plant viruses are transmitted by insect vectors , these r


Suid alphaherpesvirus 1 (SuHV-1), better known as Pseudorabies virus (PrV), an alphaherpesvirus of swine, is the causative agent of Aujeszky’s Disease. In the previous century, epidemics of Aujeszky´s disease have led to enormous economic losses in pig farms and an efficient vaccine was urgently needed. The highly attenuated “K61” strain, which was isolated after numerous serial passages on different cell cultures, published by A. Bartha in 1961, proved to be the crucial milestone in the fight against this disastrous disease. The Bartha K61 strain is still one of the best live virus vaccines in use and “celebrates” its 60th anniversary this year 2021). In the concise review by J.L. Delva et al. 021. In t summarizAlthough PrV is eradicated from domestic pigs in many Western countries, the virus is still circulating in wild boar populations coupled with infrequent infections of hunting dogs and the risk of reintroduction into farmed animals. Surveillance is thus still an important issue and discussed in three original articles ,6,7. DruPrV is not only prominent as animal pathogen but, together with herpes simplex virus 1, represents the most intensively studied and best understood alphaherpesvirus at the molecular level. Studies on PrV repeatedly pioneered research on basic principles of herpesvirus replication. Data on the complex transcriptome , the fun


To investigate the factors affecting the hospital costs in the road traffic injuries. This applied study examined the information of patients presenting to Yazd Trauma Center in 2018. The data were extracted from Comprehensive Traffic Injuries System affiliated to the center, which were described with frequency, percentage, mean, and SD, and then analyzed using independent t-test and one-way ANOVA. p<0.05). Moreover, the costs were higher in men, and in those with head and neck injuries, dead casualties, suburban high-way accidents, motor cyclists, hospital stay longer than three days, and older patients. Most injuries (%66.4) are caused by motorcycle and pertained to head region (%61.8). Some significant correlations were found among gender, type of injury, patient’s final status, site of road accident, patient’s nationality, type of vehicle used at the time of accident, length of stay , patient’s age, and hospital costs (Given the significant correlations between demographic and social variables under study, the results may be used in planning and designing strategies for controlling road traffic injuries and reducing the related hospitalization costs. Around 1.2 million road traffic deaths and between 20 and 50 million non-fatal Road Traffic Injuries (RTIs) annually occur worldwide [In 2015, 1.5 million people around the world lost their lives due to transportation trauma . It is aAccording to the published statistics, the load of traffic casualties and deaths is heavier in East Mediterranean (EMR) and North Africa compared to other regions of the world. In addition, the EMR region has the highest traffic mortality rate . Among cThese injuries and deaths have great social and economic impacts by creating a significant financial burden for individuals and families, as well as for the health care system , so thatAnnual RTI costs in developing countries like Iran is %6.64 of national gross production (39 billion US dollars) . HoweverRegarding traffic accidents, although some studies have been conducted on the prevalence, type of injuries, and severity of accidents, few studies have investigated the correlation between hospital costs of traffic casualties and variables affecting them , 16. FurThis applied study was performed in 2019. In addition, this research used a cross-sectional design to examine information on traffic accidents in Yazd Trauma Center in 2018. This center, located in the north-to-south route of the country, is known as the pole of trauma in the center and south-east of the country. Therefore, according to Iranian Insurance Company statistics, %30-40 of patients and casualties of this center come from the neighboring provinces. Besides, this trauma center is located in a province that, proportional to its population, has high rates of traffic and motor accidents, and the traffic accident mortality prevalence is reported to be 46 deaths per 100,000 cases . The required information was extracted from RTI System affiliated to this center. Previous studies obtained their information from some sources such as road traffic accidents, Legal Medicine Organization, traffic police statistics, and hospital sources. However, these valuable data seemed to be improper for performing epidemiological and statistical analyses to determine causes of road traffic accidents and to develop preventive intervention programs, may be due to inaccurate definitions of some variables and lack of registration of some necessary information in these forms such as patient’s costs, hospital indices, and multi-source information. Another weakness of the previous information sources was lack of diagnosis of accident-related disease in terms of the International Classification of Disease (ICD). Hence, this study used the comprehensive RTI registration system in which the diagnosis and type of trauma are considered based on ICD. The information extracted from this system were as follows: time of admission, time of discharge, the patient’s age, the patient’s gender, total cost of services, type of injury, the patient’s final status, day and hour of the accident occurrence, location of accident, and the patient’s nationality. Some information such as educational level and the patient’s occupation were excluded, since they were deficient. The gleaned data were coded and then imported to SPSS. Finally, the data were described with frequency, percentage, mean, and SD, and analyzed using independent t-test and one-way ANOVA. Excel was used to plot the curves. Totally, the information of 8105 RTI patients were investigated during the time interval of the study. Most of the patients (%75.7) were men and aged between 16 and 30 years old %42.1). In addition, most injuries caused by motor cycle (%66.4) occurred in the head region (%61.8). The mean hospital cost was 20.02 million Rials. The mean hospital cost was statistically significant among gender groups, type of injury, the patient’s final status, the patient’s nationality, and type of vehicle at the time of accident  and 2..1. In adDistribution of hospital costs in terms of age groups indicated that hospital costs increase with aging. Additionally, cost distribution pyramid demonstrated that hospital costs are greater in men compared to women before the age of 60 years old; however, it has been greater in women after the age of 60 years old than in men . In most countries, financing the treatment of injured people in traffic accidents is a separate fund that is paid from car insurance, public insurance or government fees , 19. ThiOur results show that the most frequently used vehicle at the time of accident was motor cycle. Correspondingly, this is consistent with other studies -23. ConsIn the present study, %99.3 of the investigated accidents occurred in urban roads; however, there was a significant difference between hospital costs and location of accident, so that accidents occurring in suburban highways imposed higher costs on victims. This is consistent with the findings of other studies , 21, 26.et al., most accidents occurred in the 20-40 years old age group [Most accidents (%75.7) occurred to men, the costs of whom were 1.5 times greater than those of women. This is consistent with the results of previous studies , 27, 28.ge group , India [ge group , and Viege group  also conge group  and Irange group  also repge group , Kenya [ge group , Spain [ge group , and Grege group , whereasge group . The mean of total costs of patients was estimated as 20.02 million Rials. This rate is less than the amounts reported in high-income countries , 36, whiRegarding the type of injury, most injuries occurred in the cephalic region so that head injuries were observed in %61.8 of the victims. The costs of injuries of hand and fingers, neck, and head were higher than those of other parts of the body. These findings are consistent with the results of previous studies performed in Iran  and otheOur findings showed that hospital costs are higher for “men, the older people, dead victims, motor cyclists, LOS more than 3 days, accidents in the suburban highways, and head and neck traumas”. There was a significant correlation between these variables and hospital costs. These results may be useful in planning and designing some strategies for controlling RTAs and diminishing hospital costs of RTI victims. Nevertheless, taking media-based preventive measures for all age groups and teenagers at schools, in order to encourage the use of helmet by motor cyclists and spread the public culture of observing the rights of passersby may be fruitful. This study explored trauma information of both ambulatory and hospitalized patients (inpatients and outpatients). This point should also be taken into account in future studies.


Merci Mon Héros (MMH) is a youth-led multi-media campaign in Francophone West Africa seeking to improve reproductive health and family planning outcomes using radio, television, social media, and community events. One component to this project is the development of a series of youth-driven videos created to encourage both youth and adults to break taboos by talking to each other about reproductive health and family planning. A costing study was conducted to capture costs associated with the design, production, and dissemination of 11 MMH videos (in French) on social media in Côte d'Ivoire and Niger. The total costs to design, produce and disseminate 11 of the campaign videos for MMH in both Côte d'Ivoire and Niger were $44,981. Unit costs were calculated using three different denominators, resulting in average unit costs of $0.16 per reach, $1.29 per engagement, and $4.27 per video view. These findings can be useful for future studies of SBC interventions using social media for framing the analysis and selecting the appropriate metrics for the denominator, as well as for budgeting and planning SBC programs using social media. Francophone West Africa continues to experience challenges in reproductive health and family planning (RH/FP), including high levels of maternal and child mortality, high unmet need for FP, and high fertility rates , 2. AdolA literature review of SBC strategies for improving RH/FP outcomes among youth ranging from adolescence to early adulthood in West Africa found evidence on the use and impact of traditional mass media interventions , but limited evidence on interventions using social media platforms . Social To fill this research gap, the United States Agency for International Development (USAID) is working in West Africa through the Breakthrough RESEARCH project to evaluate a mass multi-media campaign called Merci Mon Héros (MMH), which means “thank you, my hero” in French, being conducted through its sister project, USAID-funded Breakthrough ACTION. The MMH campaign includes radio, television, social media, and community events. One important component of MMH was the development of a youth-led campaign, beginning with a youth design challenge (YDC) at the Francophone SBC Summit in Abidjan, Côte d'Ivoire in February 2019. From this challenge evolved the “Merci Mon Héros” campaign wherein youth thank their parents, other family members, friends, teachers, and partners for communicating with them about RH and family planning (FP), and helping or supporting them when they had difficulty accessing RH/FP information or services . To helpDuring the timeframe of the analysis, 11 MMH videos were disseminated via social media  and via radio, television, and community-based events in nine countries . The videos consisted of individuals recounting their experiences when someone in their life shared information about reproductive health issues and how this information helped them better understand their health and options. This study examines the cost of designing and disseminating the MMH videos via social media, with a focus on the videos' dissemination in Niger and Côte d'Ivoire. These two countries were selected for analysis because: (1) The YDC that sparked the original MMH campaign idea was held in Côte d'Ivoire, and (2) Niger has the highest total fertility rate in the world and high levels of adolescent pregnancy , 14. TheThe primary purpose of this study was to estimate the total design, implementation, and unit costs associated with the MMH videos. Many SBC costing studies neglect estimating the design costs of SBC interventions, and even less is known regarding design costs associated with RH/FP SBC programming with and for youth . Thus, tA second objective was to examine the unit costs of MMH in Niger and Côte d'Ivoire. There are several potential program unit costs to examine with digital media, where the denominators of the unit costs  are currently being discussed and conceptualized in the literature , 17. UniThe study protocol and data collection instruments were developed by Breakthrough RESEARCH and led by Avenir Health with support from MMH key country stakeholders. The data collection questionnaire was developed based on the SBC costing guidelines . Before This costing study primarily took an economic costing approach, focusing on the resources needed to replicate the development of videos delivering SBC messaging through social media platforms in a similar setting. Included in the costs are those associated with in-kind contributions, such as donated personnel time and meeting space, which were valued at the expected costs based on information on the value of the donation provided by Breakthrough ACTION country teams. Broader societal costs were not included, such as the costs associated with the time for the intended audience to view the videos or the cost of internet access for users of social media.Subsequently, there was a workshop to strengthen capacity of the winning youth consultants, plus nearly a dozen additional young activists, journalists, and other young people working in the RH space in Francophone African countries for 2 weeks in June and July 2019. In addition, there was a 1-week refresher training on video production and dissemination for a group of paid youth consultants in February 2020. The costs associated with training and production were primarily the personnel time associated with staff and consultant trainers, travel, lodging and catering, car hires, and equipment. Equipment costs were relatively low as the videos were filmed using smart phones and open-source software; the costs of four smartphones purchased by Breakthrough ACTION and used for MMH were included in the analysis.The first MMH video was posted on Facebook in November 2019. Dissemination costs associated with the core MMH videos include personnel costs associated with social media influencers/bloggers paid to disseminate and promote the videos, social media paid advertising, and costs associated with the MMH launch in each country . While the dissemination of the videos and production of new videos continues at the time of this manuscript, the data collection for the dissemination phase reported here includes costs from November 2019 through October 2020 only.Finally, a portion of the overhead costs associated with Breakthrough ACTION's oversight of the production and dissemination of the videos were included. These include rent and support personnel  costs to support activities for the purpose of planning, coordinating, and managing youths' technical work on MMH video production.Because of the ongoing COVID-19 pandemic, the costing team worked remotely with Breakthrough ACTION headquarters and the Côte d'Ivoire and Niger teams to collect the data using the data collection instrument. Despite the communication challenges of remote data collection, in-country stakeholders' engagements commenced via a series of calls and web consultations in July 2020 to review the questionnaire and discuss the required data. The web consultations also familiarized the country teams with the data collection tool and the informed consent form. Verbal informed consent was obtained from key informants, which primarily included in-country SBC program and financial managers for Breakthrough ACTION.Starting in September 2020, the team conducted extensive in-country stakeholder engagements via a series of phone calls, emails, and web consultations to review the data provided. Where data were incomplete, the research team requested clarifications from MMH stakeholders, both at headquarters as well as in-country.Data on the reach of the MMH videos were captured via social listening reports conducted by Breakthrough RESEARCH in partnership with M&C Saatchi Intelligence . Their fCost data obtained from the data collection instruments were entered into a Microsoft Excel workbook for analysis. The total design and production costs were captured and allocated to the two study countries using the proportion of the level of effort the production team (youth consultants) and the in-country Breakthrough ACTION team spent on the conceptualization, production, and review of the videos during the analysis period. Because of the potential recall bias for respondents to accurately estimate their level of effort to support the different stages of design, production, and dissemination of MMH videos, a sensitivity analysis around this estimate was carried out by varying the level of effort in both direction by ± 5%, that is if the level of effort was estimated to be 10%, for example, the sensitivity analysis examines a range from 5 to 15%.Costs allocated to the design and production of the videos include costs associated with the YDC and trainings for creating the videos, including personnel costs for these activities. To apportion the design and production costs for Côte d'Ivoire and Niger from the total costs of the overall MMH program, the percentage of the overall design and production costs allocated to each of the two countries was based on the relative reach to these countries, based on the social listening dissemination report, with 18% in Côte d'Ivoire and 6% in Niger.The apportioned design and production costs were added to the total dissemination costs for each country. Dissemination costs include costs associated with the in-person campaign launch events, social media advertising, social influencers/bloggers, and program personnel for these activities. The launches were in-person but meant to drive interest in the broader campaign as well as the digital campaign. As such, only a portion of the launch costs were attributed to the dissemination of the videos, based on input from key staff. For costs listed in local currency, the average exchange rate for 2019/2020 of 552.47 FCFA = US$1 was used .via social media, measuring cost per exposure is particularly interesting due to the nature of social media platforms, which measures exposure differently from other SBC approaches, such as mass media and interpersonal communication. Three different program output denominators that measure exposure were used to calculate unit costs for Côte d'Ivoire and Niger:Unit costs were calculated using an ingredients-based costing approach, where all inputs were listed, their costs collected, and the contribution of these costs to the overall cost were quantified. For SBC disseminated Reach—the number of people who saw MMH posts at least once; meaning any content from the MMH page entered their screen, including when people scroll past the post quickly.Engagements—the number of times people engaged with MMH posts through reactions, comments, shares, retweets, mentions, and likes.Views—the number of times the eleven videos were viewed for at least 30 s, where each video was at least 2 min long .To calculate the number for each denominator for Côte d'Ivoire and Niger, the breakdown in Facebook users who have engaged with MMH content by location was used to best approximate country-specific denominators.The total cost to design, produce and disseminate the 11 core videos for MMH in both Côte d'Ivoire and Niger from November 2018 through October 2020 was $44,981, as shown in Unit costs are presented using three different denominators in Results of the sensitivity analysis around the respondent estimated level of effort by either a unilateral increase or decrease by 5% in the level of effort allocated to MMH videos are shown in Since the total costs of the design and production of the MMH videos were captured for all the West African core countries, the aggregate unit costs associated with design and production can also be examined using data capturing outcomes across all of the countries where MMH was disseminated. This total unit design and production cost was $57,525. When divided by the total outcome variables, the unit costs for design and production were $0.05 per reach, $0.37 per engagement, and $1.23 per view see .The use of social media for disseminating SBC messages has greatly increased in recent years, with many publications describing the uses, benefits, and limitations of social media for achieving public health objectives . Few stuApproximately one third of the costs associated with MMH was spent on design and production and two thirds on dissemination in Côte d'Ivoire and Niger. The $57,525 total spent on design and production for the region was apportioned to Niger and Côte d'Ivoire based on country specific program reach. As such, it is important to consider the total costs for design/production when considering the findings for budgeting and planning purposes, as country-specific costs can contain efficiencies from having a regional program or approach. As dissemination continues over time and as more videos are produced leveraging the ever-deepening knowledge from the original training sessions, the design and production costs per output will decrease as these initial fixed costs are spread over more persons engaging with and viewing the videos. One of the novel approaches undertaken by MMH is the collaboration with youth in the design, production, and dissemination of the MMH videos. While the counterfactual of what it would cost to design and produce MMH without the involvement with youth was not assessed in this analysis, any additional costs associated with youth-led SBC should be weighed against the economic benefits of building capacity in the next generation of SBC professionals in the region. These benefits are not captured in this analysis but have important implications for potential societal returns on investment.The unit costs for Côte d'Ivoire and Niger are very similar, with slightly higher unit costs in Niger due to the lower reach of the MMH videos in Niger compared to Côte d'Ivoire. While the design and production costs were allocated based on country reach, dissemination costs had fewer people reached, engagements, and views to be spread over. This is apparent in the higher proportion of total costs in the campaign launch costs in Niger vs. Côte d'Ivoire. The lower reach of SBC via social media in Niger is likely a function of more limited internet connectivity in Niger relative to Côte d'Ivoire and other countries in the region, where only 2% of the population is on Facebook in Niger compared to 18% in Côte d'Ivoire .Finally, the unit cost results using the three different denominators highlight the different potential approaches for assessing social media costs. The cost per reach denominator measures the cost to have the MMH videos appear on an individual's screen. By “reaching” an individual, one may scroll past the video without absorbing the content; however, by having the message on their social media feed, they have the opportunity to further engage with the content. The “engagement” denominator indicates an individual is noticing the content based on likes, shares, and comments. Finally, the “view” denominator examines how many times the video was viewed for longer than 30 s.The unit cost results indicate that the MMH videos were inexpensive to reach people with an average of $0.16 per person reached. This is comparable to the median unit costs for traditional SBC mass media campaigns per person exposed  . In the The second measure is the $1.29 average cost per “engagement,” which reflects the cost to get people to acknowledge the MMH campaign by either sharing, liking, or commenting. While “engagement” can capture a range of intensity from a simple “like” to a more participatory conversation through comments, this aggregated measure appears to be a more accurate gauge of campaign exposure. A remaining question is whether individuals who are engaging with the MMH content without fully watching the videos are still receiving the basic messages of MMH around destigmatizing and normalizing conversations about RH. Thus, future SBC impact analysis could investigate to what extent “engagement” is associated with improved knowledge, beliefs, and attitudes that the videos are addressing.Finally, the $4.27 per video view is the most expensive outcome; the unit costs are comparable to SBC unit costs for group interpersonal communication . The higWhile efforts were made to produce a comprehensive cost analysis of SBC, several limitations to this analysis should be noted. First, the cost data on the MMH videos were aggregated with costs from the broader MMH campaign and estimates of level of effort were used to isolate the costs of the MMH videos' design, production, and dissemination for the two focal countries of Côte d'Ivoire and Niger. However, these estimates may lack precision and have the potential for error. Contributing to potential imprecision is the possibility of recall bias. Staff supporting the MMH activities do not work solely on this program; respondents needed to accurately estimate their level of effort to support the design, production, and dissemination of MMH videos. Because no time-motion study was carried out to assess how staff and youth consultants spent their time on the MMH videos, a sensitivity analysis was used to explore this imprecision and found the 5% variability did not have substantial impact on the unit costs.Second, this study is novel in that no prior research was identified quantifying the unit costs associated with SBC via social media. As such, it is impossible to benchmark these results against the costs of other SBC campaigns via social media studies. The costs presented here are unique to this particular social media campaign, and a different set of videos with a different subject matter, length, or use of professionals would likely vary greatly from those presented in this analysis. While the unit costs described here can be considered against the backdrop of other SBC interventions such as “per person exposed” for mass media or “per person participating” for IPC, these unit costs are not directly comparable since they have different denominators. More research is needed on the effectiveness, costs, and cost-effectiveness of SBC using social media to appropriately benchmark MMH to similar programs. Only when several studies emerge on the costs of SBC interventions disseminated via social media, will there be enough information to properly gauge how the costs of SBC vary when using social media vs. more traditional forms of SBC dissemination.A third limitation is that this analysis is narrowed to include only the MMH videos where there is denominator data to generate unit costs. However, MMH is not exclusively a digital campaign nor are the videos the campaign's sole digital content, but rather has complementary mass media and community-based activities. While neither the cost nor the reach of these non-digital activities is included in the analysis, there are potential areas where synergies between these program components are not being captured. For example, if the videos are screened at a community event, the number of people who viewed the videos during that event are not being captured in the analysis.This study examining the cost of designing, production and disseminating the core MMH videos via social media in Niger and Côte d'Ivoire is the first to examine the unit costs of an SBC social media campaign and can be used by future research to compare findings from similar approaches, despite the above limitations. The variety of available denominators makes this a particularly interesting area for new research; thus, this study suggests that the selection of denominator is critical in determining unit cost estimates. Looking forward, SBC programmers should investigate which denominator(s)  provide the best measure to capture based on the program objectives. Additionally, future research that pairs effectiveness and cost studies on SBC via social media are essential to better understand the utility of this channel for promoting positive behavior change.The original contributions presented in the study are included in the article/supplementary material, further inquiries can be directed to the corresponding author.NB and MT conceptualized and designed the study, conducted the literature search, and wrote the original manuscript. MT, NB, EP, ZL, AD, MS, and LB had roles in the questionnaire design. MT conducted data collection virtually, data analysis, and data interpretation. EP, ZL, AD, and JH critically reviewed the data collected. All authors contributed to the writing and improvement of the data interpretation of the manuscript, read, and approved the final version.Breakthrough RESEARCH is made possible by the generous support of the American people through the United States Agency for International Development (USAID) under the terms of cooperative agreement no. AID-OAA-A-17-00018.The contents of this document are the sole responsibility of the Breakthrough RESEARCH and Population Council and do not necessarily reflect the views of USAID or the United States Government.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


Rhabdoviridae, Coronaviridae, and Paramyxoviridae. Viruses from these families have repeatedly spilled over to humans in recent decades, causing significant clinical disease and deaths. Here, metagenomic sequencing of a big brown bat (Eptesicus fuscus) submitted for rabies testing due to human exposure identified a novel paramyxovirus, Eptesicus fuscus orthorubulavirus (EfORV), in South Dakota, United States. The nearly complete 15,814-nucleotide genome shared 72% identity with that of human parainfluenza virus 4 (HPIV4), a virus that causes significant clinical disease, typically bronchiolitis and pneumonia, in children less than 2 years of age. Phylogenetic analysis confirmed a close evolutionary history between EfORV and HPIV4, reminiscent of other orthorubulaviruses with highly similar bat and mammalian species, including conspecific human and bat mumps virus, mammalian parainfluenza virus 5 and bat Alston virus, and porcine La Piedad Michoacán virus and bat Mapuera virus. These results support the idea that bats are a reservoir for diverse paramyxoviruses with closely shared evolutionary histories, compared with a number of significant human pathogens, and expand the range of bat paramyxoviruses to North America. Given the propensity of paramyxoviruses to overcome species barriers, additional surveillance and characterization of EfORV are warranted.Bats are a reservoir for many zoonotic viruses and host large numbers of genetically diverse species in the families IMPORTANCE Bats are a reservoir of large numbers of viruses. Among bat-borne zoonotic viruses, members of Coronaviridae and Paramyxoviridae have had the largest impact on human health. The repeated spillover of bat viruses to humans, often with devastating results, has led to increased surveillance and virus discovery efforts in hot spots for virus emergence, largely Asia and Africa. Apart from rabies virus, little surveillance of viruses in bats is performed in North America. Here, viral metagenomic sequencing identified a close relative to HPIV4 in a big brown bat found in a motel room in South Dakota. The virus, EfORV, was 72% identical to HPIV4, which causes clinically significant respiratory disease, mainly in children; it represents the first bat paramyxovirus identified in North America. Close genetic relationships between bat and mammalian orthorubulaviruses underscore the importance of bats as a reservoir for zoonotic viruses. Rhabdoviridae and Paramyxoviridae include bats as a reservoir for numerous species capable of zoonosis.Bats are an important reservoir for zoonotic viruses. In the nearly one century since the discovery of rabies virus in bats, a large number of clinically significant viruses have emerged from a bat reservoir , 2. RepeHenipavirus, emerged from bats in Australia and Malaysia, respectively  strain 6931. To our knowledge, this is the first bat paramyxovirus identified in North America.Despite the large amount of bat surveillance performed worldwide, comparably little surveillance has been performed in North America. In an effort to detect possible zoonotic viruses in synanthropic bats, metagenomic sequencing was performed on a tissue homogenate prepared from the viscera of a bat submitted for rabies testing. Direct fluorescent antibody testing of the brain yielded negative results for rabies virus. The bat was identified in a motel in central South Dakota and reportedly was crawling on the floor. A total of 852,138 paired-end 151-bp reads were used to assemble a 15,814-bp contig with 72.3% similarity to the orthorubulavirus HPIV4. The virus was named AAGAGGGGG-3′ (mRNA sense) located at nucleotide positions 2393 to 2401 leading to insertion of two nontemplated G residues and continuation of translation in an alternate reading frame. The remaining ORFs, for the M, F, HN, and L genes, code for 381, 544, 586, and 2,273 aa, respectively, which are 71.4, 58.2, 53.3, and 71.3% identical to those of HPIV4. An additional ORF, encoding 94 aa, was identified in the 475-nucleotide region between the M ORF and the F ORF. This ORF showed no homology to any proteins in the nonredundant database by BLASTP. Analysis of the transcriptional signals, however, failed to place the 94-aa-encoding ORF in the context of a transcriptional unit, suggesting that this gene is not expressed (Open reading frame (ORF) analysis identified canonical paramyxovirus genome organization. The partial N gene, encoding 551 amino acids (aa), was incomplete, missing approximately 6 aa based on sequence alignment with HPIV4b N, which was 74.9% identical. The nonstructural V gene was the second ORF and encodes a 218-aa protein with 37.3% identity to that of HPIV4b. The P gene, encoding a 391-aa protein with 42.7% identity to that of HPIV4b, shares its amino terminus with V, with a predicted RNA editing site of 5′-Paramyxoviridae resources page . Sequences were aligned by ClustalW, with phylogeny inferred by maximum-likelihood analysis using the best-fitting LG+G+I model of evolution; tree topology was assessed with 500 bootstrap replicates. EfORV occupied a sister clade closely related to that of HPIV4 -ATTGCTTCTCTTGCCCAAGTCCCA. Including the original sample, RNA was isolated from 90 bats that had been submitted for rabies testing. All bats were of the species Eptesicus fuscus; 84 originated from South Dakota, 5 from Minnesota, and 1 from Iowa. Eptesicus fuscus, commonly known as the big brown bat, is widespread throughout North America. The nonmigratory insectivorous bat commonly roosts in human structures, leading to frequent human exposure. The only positive sample was a submission in which EfORV was identified by sequencing (1/90 samples [1.1%]). South Dakota contains three distinct biomes, i.e., row crop and livestock agriculture in the eastern half of the state, with the western half being dominated by high plains grasslands and the coniferous Black Hills. The sample containing EfORV originated from the central high plains, from which our sample collection contained only two specimens. In addition to sampling biases, bat paramyxovirus shedding is periodic, with peak virus shedding occurring late in pregnancy and 2 months following parturition (To assess the prevalence of EfORV, a TaqMan assay was designed based on the L gene sequence, as follows: forward, 5′-turition . The batHPIV4 is a significant human pathogen. In the United States from 2011 to 2019, 2,700,135 HPIV tests were conducted for patients with respiratory disease. Five percent of those tests were positive, with a subset of 13% being positive for HPIV4 . In CanaMZ355765.The genome of EfORV strain 6931 was submitted to GenBank under accession number