The population of Europe is aging rapidly. Most community-dwelling older adults (CDOAs) want to remain in their homes, particularly those experiencing functional decline. Politicians and academics repeatedly praise technological instruments for being the preferred solution for helping older adults with deteriorating health to remain at home.This study aimed to understand the perceptions of CDOAs and their informal caregivers (ICs) and professional caregivers (PCs) about technologies that can help keep older adults at home.This qualitative study used personal interviews, focus groups, and photo-elicitation interviews to better understand the perceptions of a convenience sample of 68 CDOAs, 21 ICs, and 32 PCs.A fraction of CDOAs did not perceive technological instruments to be a very useful means of helping them remain at home. However, the ICs and PCs were more positive. The CDOAs preferred and were more willing to adopt technologies related to their mobility and safety and those that would help slow down their cognitive decline. The ICs preferred technological aids that assist in the activities of daily living as well as safety-related technologies for detecting falls and helping to locate disoriented older adults. The PCs preferred integrated communication and information systems to improve collaboration between all stakeholders, housing equipped with technologies to manage complex care, high-performance ancillary equipment to transfer people with reduced mobility, and surveillance systems to ensure safety at home.Although our study reports that CDOAs have limited interest in innovative technologies to help them remain at home, their technological skills will undoubtedly improve in the future, as will those of ICs and PCs. Technological tools will play an increasingly important role in home health care. The population of Europe is aging rapidly . Most coTechnology aimed at older adults is usually termed gerontechnology (which this paper will use as a synonym of technology). It is destined to have an important future role in providing support and solutions, monitoring health status to optimize autonomy, and improving the quality of life of CDOAs, regardless of their level of dependency ,9. GeronCornet and Carre  mentioneHow are technologies that help them remain at home perceived by CDOAs, their ICs, and PCs? More specifically, how do they rate their actual and potential utility?This study, overall, aimed to examine and understand the perceptions and utility of these technologies among physically and cognitively dependent CDOAs, their ICs, and PCs.This study was guided by the theoretical framework developed by Peek et al  and is tThis international multicenter study used a qualitative design to collect data on the perceptions of CDOAs, their ICs, and PCs using one-to-one interviews, focus groups (FGs), and photo-elicitation interviews (PEIs). Reporting on the study was based on the checklist for the explicit and comprehensive reporting of qualitative studies .The study population included CDOA men and women, aged ≥65 years, living in the French department of Haute-Savoie or the French-speaking part of Switzerland’s canton Valais, and with a medical prescription or indication for home care. The reason for examining perspectives in 2 countries was not to compare their populations but rather to explore a common issue together  and codevelop implementable recommendations for clinical practice and technology companies. ICs and PCs involved in clinical practice and employed in 1 of the 5 different community health care centers (2 in France and 3 in Switzerland) were also questioned.A similar sampling strategy was applied to CDOAs in both countries. On the basis of the comprehensive geriatric assessment database in the Autonomy Scale of Gerontology and Iso-Resource Groups (AGGIR) , researcThe AGGIR scale includes 2 types of variables: discriminative and illustrative. Discriminative variables evaluate coherence, orientation, washing, dressing, feeding, excretion, transfers, moving indoors, moving outdoors, and remote communication. These elements are assessed via a clinical evaluation carried out in the home of the CDOA by a nurse, who chooses from among 3 criteria for each variable: (1) the person can complete the activity alone, (2) the person can complete the activity partially, or (3) the person cannot complete the activity. Each criterion is also evaluated based on whether the person carries out the activity spontaneously, totally, usually, and correctly.The AGGIR scale is the official national tool in France for comprehensive geriatric assessment under home health care settings. It classifies the geriatric profiles of home-dwelling adults into 6 categories . GIRs 5 and 6 designate independent CDOAs who only receive minimal home help, such as meals, cleaning, and shopping. GIRs 3 and 4 designate CDOAs with mostly physical impairments, and GIRs 1 and 2 designate CDOAs with mostly cognitive impairments but also some physical impairments. The AGGIR tool uses clinical observations and questions ICs and older adults themselves on their need for assistance in the essential activities of daily living (ADL). Indeed, AGGIR is based on the topics in the Katz ADL scale, which also classifies CDOAs as independent or physically and/or cognitively impaired. Our study did not use the mini-mental state examination. The research team’s geriatrician (NV) clinically judged the participants’ physical and mental health statuses. In agreement with the ethical committee’s expectations concerning the use of existing clinical data, psychiatric diagnoses were not explored prospectively, as this was beyond the aim of this study. A detailed explanation of the tool is provided in All 5 centers used randomized sampling to select CDOAs in their respective AGGIR categories . This study was approved by the Human Research Ethics Committee of the Canton Vaud (CER-VD – 2017/000789), Switzerland, and the French Data Protection Authority. Data collection took place from September 2017 to June 2018. After the CDOAs, their ICs, and PCs had given their written informed consent to participate, the study nurse collected the participants’ sociodemographic, professional, and health status data (only for the CDOAs) and conducted the scheduled interviews .A randomized sample of CDOAs was selected from the 3 categories defined using the AGGIR tool .Thus, CDOAs with no limitations according to the AGGIR tool were placed in the independent group, those with physical limitations  were categorized in the physically impaired group, and those with cognitive limitations  joined the cognitively impaired group. Older adults with cognitive and physical limitations were included in the group corresponding to their most pronounced limitation.Candidates were contacted by telephone and invited to participate. Potential participants received written information about the study before any data were collected. After 48 hours to reflect, a research assistant contacted the potential participants by telephone to address any queries they may have and to obtain their verbal consent to participate in the study. On their acceptance, the assistant made an appointment with the CDOAs for providing more information about the study and to collect the signed consent to participate in the study, before the interview stage. On the day of the interview, the interviewer verified that the participants understood all the information and the implications of participation. Data collection used 10 photographs of relevant technologies and an interview guide developed from a literature review . Audio rThe CDOAs identified their significant ICs. Theoretical samples of 3 to 4 ICs per type of CDOA  were selected by each of the 5 community health care centers, and they were invited to participate in a personal interview and a PEI. Those without the capacity for discernment were excluded. Audio recordings of the interviews were made.In collaboration with the supervisors at the 5 community health care centers, 4 FGs were organized (2 in France and 2 in Switzerland), each with 7 to 8 PCs. The research team proposed meeting dates and times for the FGs. The PCs who were willing to participate signed the consent form beforehand, and audio recordings of the FG discussions were made.The research team developed and tested guides for the one-to-one interviews and PEIs for the CDOAs and ICs and for the FGs. The interview guides encouraged the participants to discuss relevant research issues by asking open-ended questions. The interviewer could reformulate, reorganize, or clarify questions to gain a deeper understanding of the participant’s answers. The guides also helped the participants to better identify practical technologies that might improve health care ,31. DataDescriptive statistics were compiled to describe the participants’ sociodemographic, health, and professional characteristics as well as the technologies selected. Statistical analyses were performed using the IBM Statistical Package for the Social Sciences 25.0 . PersonaThe study sample was composed of 68 CDOAs, 21 ICs, and 32 PCs. The distribution between the French and Swiss centers was 40 and 28 CDOAs, 13 and 9 ICs, and 16 and 16 PCs, respectively.Of the 68 CDOAs, almost three-quarters were women , with an average age of 82.3 years ; approximately two-thirds  lived in urban areas. The average age of the 21 ICs was 68.4 years , with 16 women and 5 men. Most were retired (n=13), lived in urban settings (n=13), and took care of a loved one with a cognitive impairment (n=12) who required partial assistance with the ADL n=6; .A total of 4 FGs of 8 participants each brought together 32 PCs, all employed at community health care centers. Their average age was 46.7 years  and most were female. Their varied professional roles included physicians, nurses, social workers, nursing assistants, care assistants, and occupational therapists .We recorded a total of 40 hours and 53 min of personal interviews and PEIs with CDOAs  and 12 hours and 25 min with their ICs . We recorded a further 4 hours and 50 min of interviews with the 4 FGs .In light of the theoretical framework adopted , we applCDOAs perceived technology as something that could be useful, especially for other older adults with health problems, but they rarely saw any sense in using them themselves, except when they had significant difficulties with their ADLs. CDOA 6 said that technology was:For people who can’t get anything done alone.CDOA 63 thought it might be needed in future:Not for the moment, anyway. Maybe in a few years, yes, let’s say, if I have problems in a few years, because I have...well, because of my age!”CDOA 53 had negative perceptions:Because all these...these modern techniques, and everything, don’t interest me much. Because it doesn’t match my past life at all.CDOA 73, however, held an opposing view:But if something can make life easier, why not? At my age, now’s the time to try.The concept of technology, in general, was often misunderstood. CDOA 75 exemplified this:Technologies? What do you mean by technology?Some CDOAs stated that technology could be useful but not in their current situation. CDOA 9 stated that they did not see any need for it in their current situation:It works automatically, huh? Of course, but for the moment, it’s not for me.CDOA 13 thought that some devices might be useful to them if they had more severe health problems:I think that I’d have to be really bad, huh, to put this thing on.The (positive or negative) interest in technologies was also often stated by this group. CDOA 9’s positive perceptions of technology were typical:Well, I think it’s...it’s 2017; I think it’s a necessity, isn’t it?CDOA 85 spoke out against technology:And then, it’s no good at all. Sometimes it’s completely harmful, like when things are made available to people, like me, who aren’t always able to understand.technology, without context, was not really understood. As CDOA 82 said:Among CDOAs with mainly cognitive impairments, the word I don’t know what that means.independent group mainly spoke about how technology could be useful for others. CDOA 7 gave their opinion:The relatively Well, it’s good for people who don’t have many visitors or people who don’t get any help from their families. And I’m not one of them. So then, no.technology itself, as in the other groups, came up regularly. CDOA 10 expressed this as a lack of knowledge:Misunderstandings about the term and the meaning of I don’t know which technologies, that’s the problem. That’s because I don’t understand everything...I don’t get it...because, well, I know there are a lot of things now, but I don’t know everything about them.Positive and negative assumptions appeared in similar proportions among the independent group, as they did for the other 2 groups. For example, CDOA 72 gave their opinion on the internet:Today, what I don’t agree with is the internet. It’s going to destroy the planet. That’s it, one way or another because it is so dangerous, especially because of those who know it very well, they could flatten the whole world. So, I’m really not interested in the internet; so I’m not interested in computers either.CDOA 83 shared their positive perception:If it’s easier for us, if it can help us out, why not, eh?Some ICs considered technologies potentially useful in helping CDOAs remain at home, whereas others considered them as indispensable. Their perceptions on the usefulness of a technology seemed to depend mainly on their current state of health, their own openness to technologies, and the costs involved. Different ICs expressed these aspects as follows:The thing that might encourage me is if it, err, her case worsened.IC 38She has already been offered them, but she doesn’t want them.IC 56Yes, change. I think if we change things, I think it would be complicated for her.IC 38Oh, well, that must be very expensive.IC 43PCs perceived technology as useful, but they specified that there was the risk of creating distance in their relationship with the persons in their care and that technology must complete and complement a PC’s work, not replace it:It can only be complementary. At some point, it makes sense if a real person has to be involved.FG 2Moreover, the indication chosen for the use of a specific technology should not only reassure ICs:What worries me is situations like this, where the children have even put cameras in the bedroom, not to monitor their parents, but rather to reassure themselves.FG 2Technologies that could be introduced to enhance the daily lives of CDOAs include furniture with home automation technologies, suitably adapted public transport, and an elderly friendly urban architecture in environments that facilitate transport mobility:We often have patients who have to take taxis or be driven to hospital or to a medical consultation in town...It’s a real transportation problem.FG 2Fifteen years ago, they were all at home, they didn’t move. Now you see some, I don’t know how many, in the city center, using their rollators. It’s pretty impressive. Soon, they’ll invent rollator parking lots.FG 1Urban living could also be rethought, perhaps by setting up social centers for CDOAs, on the same principle as youth centers:Architecturally, it would have to be equipped with ramps for the handicapped, for the rooms where they can meet up. As we do for youth centers, why not build a place for older people with physical difficulties?FG 1PCs highlighted their lack of knowledge about the technological devices available on the market and their lack of training on how to pass on skills to CDOAs and ICs:I don’t have much knowledge about this. It’s true and it makes sense that some training on this wouldn’t be bad.FG 2physical and cognitive impairments, encouragement to use technology by children and grandchildren was mentioned as an important strategy. Family members often gift devices that they no longer use to their older adult relatives. Children choosing devices, such as remote alarms, was also a facilitator of technology use. CDOAs mentioned that receiving financial aid for the acquisition of a device was a facilitator, as was receiving recommendations from PCs. CDOA 35 exemplified the assistance received from PCs:Encouragement from children and grandchildren, financial help for purchasing technology, and increasing feelings of safety were the main strategies identified by CDOAs themselves for improving the ease of use among their peers. For the groups with mainly There is a home for the blind; they proposed it to me. And they sold me the [talking] watch.CDOA 76 stated:Yeah, that’s right. So, my granddaughter has a tablet, and my son put my number into it.independent CDOAs, financial support for buying a device and receiving help on usage were the main facilitators of their use. CDOA 84 explained their situation:Several facilitators to improve beliefs about and attitudes toward technology emerged during the interviews, one being that a device can improve feelings of safety. Financial assistance can be considered a facilitator because it influences people’s willingness to acquire a device. Indeed, for the group of Listen, my son gave me this mobile; I didn’t want it. And he said to me, “It’ll be easy.” Anyway, there you go.CDOA 83 said:Someone should come and explain and turn it on and say “You have to do this and do that.”feel safer was the main facilitator of use among this group. CDOA 84 explained this in relation to their personal situation:Again, with regard to beliefs and attitudes, a technology that made participants And then I was unsure, and I was thinking about it, but as I said, I was putting it off and putting it off. And then there’s the personal side, you see? I have great neighbors, but the young ones work, and the old ones are no stronger than me, right? So, I said, “Oh, well!” and then it’s a bit of a safety thing, even though, as I said, I hope I’ll never need it.Devices’ properties, such as ease of use, also influenced CDOAs. CDOA 10 described using a stair-lift:Yes, usage. You press a button; there’s nothing complicated, eh? There are knobs, one at the top, one at the bottom, so it depends on where you are. Well, you grab the knob. And then, if you go up the stairs, you use the stair-lift device.Some ICs supported the use of technologies if they found them useful and easy to use. As IC 43 said:If it is a technology that can help me, I will accept it. If it can lighten the burden of helping her.Surprisingly, other ICs would only introduce certain technologies if the CDOA’s state of health deteriorated. Nevertheless, ICs often felt that they lacked the knowledge to use technologies properly. As IC 71 said:I like the technological side, but I just have one problem with it. It’s clear that people who weren’t born in the technological age, or who haven’t got themselves up to date by 50 or thereabouts (are going to have trouble). I even find that I’m often overwhelmed.ICs also noted their relatives’ lack of motivation about the introduction of new technologies:Ah, well, it’s certainly a very good thing, but she still has to want to do it. She’s not on board with all that’s modern. So, she’ll say no to thatIC 43The ease of use of technologies was overrided, as perceived by the ICs, by the fear of making mistakes when using them or of infringing on the CDOA’s privacy and autonomy.PCs mentioned that the context of technology implementation was important in its ease of use:Older adults just need to be interested in new technologies. They are pushed towards it by their children—by their grandchildren, especially.FG 2Sometimes a neighbor will say, “You know, I’ve got a rollator, and it goes great,” and that’s it. It’s not some young woman who comes in and says, “This is what you need.”FG 1Indeed, technologies were accepted more readily if relatives and ICs were involved:Sometimes they actually need their family’s support in order to be reassured.FG 1Ease of use also seemed important:There are some things that could improve autonomy as long as they remain simple and easy for people to use.FG 2According to PCs, the CDOAs who had either found a technology useful or had used technology in their working lives were more willing to use the proposed technologies:Some CDOAs are familiar with Skype, for example, using it with grandchildren abroad or even in another city.FG 3The elderly population that needs these aids now, is often a population that is unaware of the digital revolution.FG 4Although the point of view of individuals and the context in which technologies are implemented tend to be the factors that promote technology use among CDOAs, the cost involved in acquiring different technologies and the different steps required to obtain them can be seen as barriers:And I find that they’re always scared that “It’s going to be expensive; I can’t.” Sometimes they don’t realize the means they still have, whether financial or not.FG 4The emotions associated with technology use can also be a barrier when they generate stress and fear:The more things or equipment involved, the greater the source of stress, for the patient and the family.FG 4You get to a certain point in the aging process where people are not necessarily very accepting of their loss of autonomy. And consequently, in accepting any assistance that might be provided to them.FG 4For all groups of CDOAs, the acceptability of using technologies was influenced by the self-developed strategy for achieving this purpose, difficulties in using the device, and the technology’s cost. As an example of avoiding technology use, several CDOAs said that a simple bedside lamp gave out sufficient light to get to the toilet. The CDOAs found no reason, for example, to replace the home helps who came to clean their home with a robot vacuum cleaner. Help provided directly by humans was the preferred option for many CDOAs. CDOA 13’s thoughts on electronic pill dispensers began with a deep breath:I’m telling you, I don’t feel the need for one. Oh! If I have something special, my daughter still comes, three times a week, and she is a nurse.physical impairments did not find some device properties to be user-friendly, and multiple participants considered this to be the main obstacle. CDOA 35 complained about their tablet:The CDOAs with mainly Because handling this device is very complicated for me, you see? I have to turn it all over the place to find out what is going on...No, I’m just not interested.Another barrier was the cost of acquiring a new technology, as explained by CDOA 6:Oh, yes, because I can’t say I’m going to buy anything, in addition to what we have to buy every day, because we can’t afford it.cognitive impairments, the main barrier was the difficulty in using technologies. This was reflected by CDOA 20:For the CDOA group with mainly So, to start with, I don’t want a computer. That’s out of the question; I don’t want a computer. Well because I wouldn’t know how to use it. That’s why.In several interviews, the group members were found to prefer human contact over technological assistance. CDOA 76 expressed doubts about robots:Because there’s a person, the nurse, working; a robot isn’t human, whereas a nurse can talk, say something nice, give you a little smile. But a robot’s just ”Crash! Bang!”’Financial hardship was also consistently mentioned as a barrier to technology use. CDOA 76 said:Yeah, robots. Yeah, but you need the means to buy one.independent CDOA group, financial and usage difficulties were also the most frequently mentioned obstacles. Regarding financial aspects, CDOA 10 said:For the Well, let’s say, it’s a good thing. It’s good, but it must have a cost. Well, yes because...I still have to pay for the nursing home; don’t forget that. My husband’s pension, unfortunately, only pays for half of it.Regarding usage difficulties, CDOA 10 described their tablet:Right now, I do not know how to use it. My daughter, she’s got one. My grandsons have one each, but so far I’m not interested.Technologies designed to promote older adults’ home support, such as fall detectors, GPS, and brain training, are the ones that are most frequently accepted by CDOAs. CDOA 73 explained their acceptance:But we don’t know if I’ll ever be able to...it’s because I’m a little scared, that’s why I don’t go out alone. I’m a little scared to cross the street. So that’s why it’s really the fall detector that would suit me best.Regarding brain training, CDOA 63 explained:Well, because by...improving your powers of concentration, developing your vocabulary, anyway because of the propositions that are there, I think it’s good. That’s why.Perceptions about the acceptability of technology use were significantly influenced by safety, ease of use, keeping CDOAs autonomous, and the costs of devices.ICs explained how the acceptability of technologies was related to the safety of their loved ones. This result can be explained by the fact that of the 21 ICs interviewed, 12 were relatives of CDOAs with mainly cognitive impairments. Thus, the technologies chosen by the ICs were very similar to those chosen by the group of mainly cognitively impaired older adults. Opinions of the ICs regarding the acceptability of the technologies presented during the PEI interviews are as follows. For the light path selected by several ICs, the caregivers said:Yes, I think that is really good. Well because my mum gets up at night, she can’t find the light.IC 42That’s right, because I see he gets lost going to the toilet.IC 44Concerning the fall detector, accepted by about one-third of the ICs, they said:Yes, that could be the most useful there. [...] in the future there could be others [devices].IC 43Maybe rather the fall detector, I’d say.IC 55The ICs were more reluctant about the electronic pill dispenser:Ah, well [...], it could be useful because I want him to have a pillbox; the nurses come to put the pills in. But if he doesn’t think about it—if I don’t think about it, he doesn’t think about it—then it’s not much use.IC 29Yeah, it’s not bad, but I think it’s pretty expensive for how much we use it. As long as we’re here, well, they’re taken at mealtimes, so I’m here [laughs].IC 40[…] In my mother’s case [...] she’d be unable to take her medication alone.IC 42The ICs had diverse opinions about robot vacuum cleaners:For me, these robots are good. You shouldn’t have anything on the ground [laughs] [...] You shouldn’t; it doesn’t go in corners, it’s round [laughs].IC 29Well, it depends on how much it costs, huh?IC 38Not essential, in my opinion.IC 40[….] I think not. No, it might be more dangerous than useful.IC 42As for the opinions on the service robot, the ICs said:Yes, listen, yes, it’s good, but in the end, a human being is still better.IC 23[…] I find it rather stressful.IC 29Opinions on the usefulness of a GPS bracelet could be summed up by this statement:That, I, I’m for that. If she ever decided to go somewhere, it would let her know where she was...it would let us know where she was.IC 38Only a fraction of the ICs found the touchpad to be an interesting technology. There were different opinions on this:No, so my mom, my mom... she’s not interested.IC 23It might make things easier for him, but since he was not very enthusiastic, he quickly lost patience.IC 29Uh, no [laughs], there. Uh, I think it’s impossible. Because even a mobile phone would be complicated to use, so...He still manages to work the TV, but that’s all.IC 55Among the ICs interviewed, the idea of a social network for ICs raised little interest:I think it’s good… For the moment, I don’t need it, because...Well, I have a good rapport with the home help; it’s going well.IC 23I’m not…I’ve never been a person like that, not very communicative and that...So, everyone has to say what they do, what they’ve got...It’s not...No, not for me. But I’m not saying that it might not be a help to a lot of people.IC 29Oh! It’s not bad. It could...for someone who is alone, be a help, yes.IC 40Brain training technology was selected by only one IC, but the quoted text below illustrates the difficulties in using this technology:She can’t do it alone either. Well, memory exercises would be great, but maybe someone would have to help because of her not being able to use a tablet.IC 42To the ICs, activity sensors were not a promising technology to strengthen home support:Ah! Well, for people who live alone, that’s very good.IC 23Oh, yeah, sometimes he hasn’t closed [the fridge door] properly. It’s happened sometimes, when it didn’t shut properly.IC 29[…] she’ll say to me: “You don’t think I’m crazy yet, do you?” All this scares me.IC 38Yes. Yes, well that could be, err, that could be good, indeed. Yeah because in half an hour anything can happen, eh?IC 42[…] I wouldn’t even trust the sensor [...].IC 44[…] it’s true that these are technologies that are great for those who are not safe, but who are in their right mind, and everything, who don’t have all their heads and bodies; and who may be putting themselves in danger, uh, about that.IC 55Among PCs, perceptions about the acceptability of technology differed according to the constraints that technologies impose on the CDOAs. The more technologies focus on monitoring a CDOA’s behavior, the more ethical questions they raise. On the contrary, the more they increase a CDOA’s autonomy and safety and decrease the burden on ICs, the more the PCs promoted them. Opinions on the acceptability of some of the technologies proposed during the PEIs were as follows. The light path technology was broadly well-received by the PCs, and the following statement illustrates their favorable opinion of this technology:A technology that should be introduced and available for all older people.FG 1Technologies that increase safety, such as the fall detector, were mostly preferred by the PCs:[…] if it’s good, it can be very useful.FG 2The PCs showed little interest in the robot vacuum cleaner and the service robot:[…] There’s a risk of falling over the vacuum cleaner when it’s running...FG 4[…] It’s interesting if it assists caregivers [gives them time to do another activity while the robot is working].FG 1There's a risk of replacing humans […].FG 3Several PCs had questions regarding the GPS bracelet technology:[…] it’s not relevant if its only purpose is to calm the family […] .FG 3[…] there are ethical problems if it’s used as a means to monitor the person […].FG 3The touch tablet and brain training technologies were rarely chosen by the PCs. Their reluctance was expressed as follows:You have to accompany the arrival of the tablet in the person’s environment and ensure that the person is interested in using it….FG 4It might be interesting to couple a tablet to the remote alarm [to enable visualizing the person’s condition if they can’t or can no longer express themselves because of cognitive disorders]...FG 4[…] difficult to offer if the person does not know how to use a tablet…FG 4...can be a prevention tool…FG 3Although some PCs selected the social network and activity sensor technologies, there were mixed opinions:[Social networking] technology is not necessarily appropriate if the caregiver does not understand those technologies; it’s rather aimed at young caregivers [...].FG 2[…] it can make caregivers feel supported [...].FG 2Concerning activity sensors, this response was obtained:[...] Pose an ethical dilemma [...]; [...] culture can be an obstacle to the use of this technology; [...] interesting technology, and install the sensors only as needed . It can be a complementary tool in food monitoring if this problem has been previously identified [...]FG 2When choosing the most useful technology overall, the PCs differed according to their profession. Physicians, nurses, and occupational therapists chose the fall detector, whereas nursing assistants, social workers, and direct community health providers chose social network. To the best of our knowledge, this is the first research in the field to have combined personal interviews, FGs, and PEIs in the same study. Our results show that technologies are not yet well integrated into the daily care of frail or cognitively impaired CDOAs. Although technology can certainly offer solutions to some of the specific impairments typical among CDOAs, ICs and PCs still have little knowledge about their existence and how they should use them in everyday practice. The heterogeneity of participants’ profiles made it difficult to compare this study with others involving more homogeneous samples ,39.Our results indicate that most participants had both positive and negative attitudes toward technology. Positive attitudes were most often related to technologies that ensured continued mobility at a lower cost and with advanced functions (a GPS bracelet). Negative attitudes of CDOAs were often associated with the risks of becoming technology–dependent and the risk of fewer or poorer interactions with ICs and PCs, the realization that they were no longer able to stay at home, potential social problems , and the financial costs of technology use. Furthermore, the findings suggest that ICs had more positive attitudes toward assisting technology and an increased acceptance of technology than the CDOAs themselves. This could be explained by the fact that the average participating IC was 14 years younger than their CDOA and thus had more contact with technology in their daily life . NeverthTechnology use was often conditioned by the multiple expected outcomes CDOAs had with regard to remaining at home, risk control by ICs and PCs, and trying to avoid hospitalization or institutionalization. In contrast, the nonuse of technology was linked to the participants’ use of their personal and individual capacities, their health, their physical and intellectual functional capacities, and the environmental barriers they encountered . To encourage the participants to adopt technology, our findings suggest that the potential barriers to this should first be removed at the individual level , then at the technological level , and finally at the environmental level (life context). These results were consistent with the research conducted by Chen and Chan .Results from the PC FGs revealed that they had mixed attitudes toward technology, although most were interested in specific technologies for everyday use. They highlighted that each care situation was unique, uncertain, and often complex. They were particularly interested in the development of new technologies to help with the ADL. Finally, barriers to the use of technology by PCs themselves included lack of interest, a need for training, the architectural inadequacies of house design, cost issues, and their fears of being replaced by that technology and of the risks of dehumanizing care relationships with CDOAs. Our results highlight the need to pay more attention to adapting technology to users’ personal preferences, while focusing on technologies that provide solutions to individual problems. However, to date, there is little evidence to evaluate the usefulness or effectiveness of certain technologies in home care and notably to avoid hospitalization . Causal Technologies in home health care settings must also be adapted to the needs and concerns of PCs . In lineThe development of technologies to strengthen home support and prevent the loss of autonomy is a demographic, human, social, and economic imperative; it must be accompanied by the development of multidisciplinary skills . AcademiThe significant sample of CDOAs, ICs, and PCs involved in this study suggests that our results may be transferable to other regions of Western Europe. The PEIs made it possible to clarify perceptions and specify acceptable technologies. Another strength of this study was its use of a novel combination of methodological approaches, via one-to-one interviews, FGs, and PEIs. This approach illustrated which technologies might be acceptable and useful to CDOAs, ICs, and PCs in the context of home care support. Our research population’s heterogeneity did not permit a thorough comparison with other studies in the literature as they involved more homogeneous samples -20. In aDespite the omnipresence of technology in modern society, research into perceptions about it and its use to improve the daily lives of frail and cognitively impaired CDOAs is poor compared with research into its impact on younger populations. Despite some evidence that technologies promote independence among CDOAs, they are often underused in daily home health care. With this in mind, this study explored the perceptions about technology’s place in home health care among CDOAs, ICs, and PCs. Our findings showed that although many technologies were available to support independent living for CDOAs, they were significantly underused. Our results also affirmed that efforts should be made to adapt technologies to the needs of CDOAs, their ICs, and PCs. It seems important to continue searching for empirical evidence of the relevance and effectiveness of new technologies. This will help to specify the most important areas requiring intervention and indications for technology use among different profiles of CDOAs, their ICs, and PCs. Technology will also help to optimize the management of CDOAs’ health problems and slow their loss of autonomy, both of which will strengthen home health care. Progress toward this goal will only be achieved through close cooperation between technical experts, home health care experts, and the end users—CDOAs, ICs, and PCs—who need the appropriate technological tools to meet their needs and expectations. Finally, as the mean age of the population is rising, the proportion of older adults with an interest in technology and with technological skills will increase. The findings in this study will enable future CDOAs to clearly express the advantages and limitations of the technologies in their lives. The current situation is therefore very fluid, and research will have to adjust to this dynamic process. Although there is no doubt that technologies will play an increasingly important role in health care services for CDOAs and in the work of their ICs and PCs, it is more difficult to predict which types of interventions may develop. Establishing convincing results based on robust scientific evidence will be difficult, but clinical research will play a key role.


Viruses on “Plant Virus Pathogenesis and Disease Control” give new, important information addressing different aspects of this topic.Plant viruses are emerging and re-emerging to cause important diseases in many plants that humans grow for food and/or fiber, and sustainable, effective strategies for controlling many plant virus diseases remain unavailable. However, cutting-edge technological approaches in cell and molecular biology and rapidly accumulating virus sequence datasets are allowing us to gain new insights into mechanisms involved in pathogenesis and how interactions between plants and viruses affect disease development. In some cases, these offer opportunities for novel approaches to target plant viruses and help control the diseases they cause. Seven original articles in this Special Issue of Solanum lycopersicum) in several parts of the world. Klap and colleagues show further complexity associated with TBRFV-induced disease. They show that when TBRFV co-infects plants with the potexvirus, pepino mosaic virus (PepMV), more severe symptoms develop [Tomato brown rugose fruit virus (TBRFV) is a recently described and rapidly spreading tobamovirus affecting tomatoes ( develop . PepMV cGlycine max) are a globally important crop plant and soybean mosaic virus (SMV) occurs wherever soybeans are grown, in part due to its seed transmissibility. Bao and colleagues developed a GFP-recombinant SMV system to assess SMV-soybean interactions [Soybeans  technology to assess the role of microRNA nbe-miR1919c-5p in infections of N. benthamiana plants by tobacco curly shoot virus (TbCSV) and its betasatellite (TbCSB) DNA [Three groups addressed the roles of non-coding RNAs in plant virus infections ,6,7. LiuThese papers are representative of the approaches and opportunities that are available now to gain a better fundamental understanding of plant virus-induced pathogenesis. Obtaining a more fundamental understanding of plant-virus interactions is necessary if we are going to develop efficacious, environmentally sound approaches to control plant diseases caused by plant viruses. We now have fantastic new tools including NGS and bioinformatics approaches to look at virus diversity and host responses to virus infection at the transcriptome but also small and large non-coding RNA levels, not only in different plant genotypes but also in different plant tissues. However, the diversity of papers here also shows that we cannot ignore virus-host biology, and we must consider biology to help us accurately interpret and use what we are finding with modern technologies.


Helicoverpa armigera was first detected in Brazil in 2013 and generated a wave of speculations about the possibility of hybridization with the native sister taxon Helicoverpa zea. In the present study, we used genome-wide single nucleotide polymorphisms from field-collected individuals to estimate hybridization between H. armigera and H. zea in different Brazilian agricultural landscapes.Invasion of organisms into new ecosystems is increasingly common, due to the global trade in commodities. One of the most complex post-invasion scenarios occurs when an invasive species is related to a native pest, and even more so when they can hybridize and produce fertile progeny. The global pest H. armigera incursions is reflected in the structured and inbred pattern of genetic diversity.The frequency of hybridization varied from 15 to 30% depending on the statistical analyses. These methods showed more congruence in estimating that hybrids contained approximately 10% mixed ancestry (i.e. introgression) from either species. Hybridization also varied considerably depending on the geographic locations where the sample was collected, forming a ‘mosaic’ hybrid zone where introgression may be facilitated by environmental and landscape variables. Both landscape composition and bioclimatic variables indicated that maize and soybean cropland are the main factors responsible for high levels of introgression in agricultural landscapes. The impact of multiple H. armigera and H. zea in agricultural areas. Continuous monitoring of the hybridization process in the field is necessary, since agricultural expansion, climatic fluctuations, changing composition of crop species and varieties, and dynamic planting seasons are some factors in South America that could cause a sudden alteration in the introgression rate between Helicoverpa species. Introgression between invasive and native pests can dramatically impact the evolution of host ranges and resistance management.Our data showed that the landscape composition and bioclimatic variables influence the introgression rate between  An invasive pest can cause adverse effects of various degrees of severity, as high adaptation potentials and dispersal can cause dramatic costs to ecosystem services and agricultural production –4. ManagHelicoverpa armigera (Lepidoptera: Noctuidae) is native to the Old World  and is one of the most important pests worldwide  ; zea)], while NewHybrids identified AGOSA as the most extensively “hybridized” . The NewHybrids approach detected fewer putative hybrids in our samples than structure analysis, and successfully flagged one putative F1 hybrid in AGOSA  , based on the locations sampled. The main direction of interspecific gene flow seemed to be from H. zea to H. armigera, affecting insects from APRLO, AMCV, and AGOSA   Fig. a. The moH. armigera   Fig. b. The cuχ2 = 7.69, p = 0.02), climate-only model , or the landscape-only model .When we compared different model sets that contained explanatory variables , we determined that the full model best explained the observed variance (AIC = 94.7), compared to the naïve model  into the pure lines, creating an adaptive bridge between the two species. Multiple hybridization events can enhance the fitness performance of the two species involved, even when the rate of hybridization is relatively limited [We hypothesized that relatively few hybridization event occur but involve introgression of large genomic areas . Our dat limited . Under t limited .H. zea [H. armigera in Brazil [H. zea. This introgression provides compelling evidence for the potential adaptive advantage of hybridization, especially in agricultural systems. The implications can extend beyond insecticide resistance and affect other traits such as host range. For example, H. zea has lost a significant number of detoxification genes and gustatory receptors due to genetic drift, and might be ‘re-acquiring’ some of the ‘lost’ genes from H. armigera [H. armigera spreads and overlaps with another Helicoverpa pests in Argentina (H. gelotopoeon) and with H. zea populations from North America [Hybridization can also have profound repercussions for the native species, as demonstrated by the recent detection of the CYP337B3v2 resistance gene in H. zea . This ubn Brazil , and nowarmigera . The sce America .H. amigera ecotypes or to fusion into a single panmictic population. However, in order to determine the evolutionary trajectory of the two genetic groups, hybrid viability must be determined. If hybrid viability proves to be limited, then maintenance of the two species is the most likely scenario. On the other hand, if hybrid fitness exceeds that of the purebred lines, then the species are expected to more closely resemble one another. Therefore, while hybrids may perform poorly in a laboratory setting, some hybrids may have beneficial qualities that increase fitness in a complex mosaic of agroecosystem. Our data indicate that hybrids are present in natural populations; whether or not the level of hybridization will increase or not needs further investigation.In relation to the viability of hybrids, the interaction between genetic and environmental factors has shaped and will continue to shape the distribution of genetic diversity, leading either to the development of H. armigera invasion in South America are the high mitochondrial haplotype diversity , the genetic similarity among distant parts of recently colonized areas, and the differences in regional dynamics  [H. amigera sampling locations, which might suggest some level of genetic structure. In addition, the higher values of FST among H. armigera populations can be explained by hybridization with H. zea, the presence of multiple H. armigera lineages, genetic drift , and differences in local dynamics . Many questions remain regarding how the various invasion events occurred in Brazil, such as if the invasive specimens originated from a pool of founders of mixed ancestry or if the admixture occurred upon arrival. The patterns of genetic substructure and intra-species hybridization within H. armigera populations captured by our data may suggest the presence of multiple H. armigera lineages that are partially admixed. In contrast, the FST values for H. zea do not suggest a high degree of genetic differentiation, supporting the evidence for panmixia, at least within Brazil [The three major features of the osition) , 32, 33.n Brazil , 34, 35.H. armigera populations has always been a contentious topic of debate [H. armigera seem to be experiencing extensive gene flow in other parts of the world [H. armigera may not reach a level of panmixia like H. zea [H. armigera’s population structure, which will improve the predictions how resistance might spread.The genetic structure of f debate –38, wherf debate . Populathe world . We can,e H. zea , 38. Con, up to 90%) [Similar to other previous research using SNP data, we have also detected a substantial rate of missing data caused by the interruption of the recognition site of restriction enzymes . The hig to 90%) , 39. NonH. armigera and H. zea hybridization instead formed a “mosaic hybrid zone” where the patchy hybridization hotspots have no apparent spatial pattern [H. zea is predominantly associated with maize, whereas H. armigera is often found on soybean and cotton. The mosaic configuration of the agricultural landscapes and the intensity of Brazilian farming (two to three crop seasons in a year) facilitate the simultaneous production or succession of suitable host types in the same area. Our study provided evidence that rotation among crops can be particularly problematic and increase the probability of hybridization. Furthermore, both species have resistance to management practices: H. armigera is resistant to commonly used pyrethroids and H. zea is resistant to the Cry1Ac Bt-protein present in some transgenic soybean. Therefore, we can expect to see the first signs of insecticide or Bt resistance caused by introgressions and host-changing behavior in areas with extensive production of maize and soybean. In areas where these crops do not coexist, hybridization levels are likely extremely low, indicating that the appropriate choice of crops to rotate and the use of polycultures are essential for preventing and managing hybridization in Helicoverpa.Rather than forming parallel clines where admixture can be easily recognized,  pattern , 41. A cH. armigera and H. zea in Brazil. According to the different methods of inference, hybridization between the two species ranged from 15 to 30% among Brazilian locations. No significant asymmetry in hybridization between the two species was detected, but the probability of hybridization and the extent of the introgression were significantly affected by the environmental conditions, including climate and landscape composition. Insects from locations where maize and soybean were present tended to show high levels of hybridization. The most concerning finding is the continuous exchange of adaptative genetic variation that will likely affect the host range and insecticide resistance. If hybridization continues and increases it will likely complicate the management of these pests and further threaten crop production in Brazil. Continuous monitoring of the hybridization process is necessary because of the expansion of agricultural areas, climatic changes, the composition of crop species and varieties, and the planting seasons in South America. These constantly changing factors could lead to sudden changes in the rate of introgression between these Helicoverpa species, and strongly impact on the host range and resistance management.In summary, we have found strong evidence for hybridization between Helicoverpa species were collected from 13 different Brazilian locations by active searching on host plants. The sampling included the most important soybean, cotton, and maize-producing areas in Brazil during the 2015 crop season. Detailed information about the host plant, collection date, and geographic coordinates is presented in Table Larvae of both BstZ17I [Total DNA was extracted from each specimen following an adapted protocol based on the CTAB method . After tBstZ17I .Helicoverpa species (53 H. zea and 119 H. armigera) were selected to generate two GBS libraries containing ~ 86 insects each [MSeI). Last, we used HiSeq 2500 to sequence the pair-end libraries, which were prepared and sequenced at the Molecular & Cellular Imaging Center Genomics Facility at the Ohio State University . Raw fasta files of Illumina sequences were included in the SRA-NCBI repository (PRJNA615801).A total of 172 samples of cts each . Before process-radtags implemented in STACKS 2.2 [de-novo approach to assemble loci has been extensively used in non-model system and when there is no reference genome available; this strategy is also more appropriate when the percentage of alignment is low. We ran the de-novo pipeline using all default parameters, closely following the method described by Anderson et al. [tsv2bam and assembled into contigs using gstacks. In the last step, we generated statistical summaries and Treemix analysis using the population module, allowing a minimum of 5% individuals required within groups and 100% between groups, excluding SNPs with less than 5% frequency, using one random SNP per RAD locus. Due to the great divergence between groups and the possibility of a high degree of variation within groups caused by hybridization, filtering parameters were relaxed, allowing an overall 20% presence of SNPs . We conducted preliminary tests to maximize data retention while minimizing the rates of missing data in both species. The impact of hybridization varies in different parts of the genome, as previous studies have shown [Raw-sequence reads were demultiplexed using ACKS 2.2 , 46. Rean et al. . After rn et al.  providedve shown . Thus, ave shown , 49.q), representing the proportion of the genotypes originating from cluster categories (K), was later used to infer the putative degree of introgression in each sample. We used individual estimates of the introgression of insects collected at different locations as a dependent variable in models to explain possible causes of the observed differences.Species were identified using the collection information, including the host plant, morphological characters, and mtDNA genotyping, followed by the analysis with SNPs. Bayesian clustering methods implemented in STRUCTURE 2.3.4 and NewHybrids 1.1 were used to identify putative hybrids and to estimate proportions of nuclear admixture and patterns of introgression –52. For H. armigera [H. armigera in Brazil. We ran the STRUCTURE analysis for a range of K values (K = 1–10), and subsequently used the Evanno method implemented in STRUCTURE HARVESTER 0.6.93 to test for the most likely number of K [n = 10). Replicates of each K value were aligned and averaged in CLUMPP 1.1.2 [First, we assumed K = 2, because two gene pools could potentially contribute to the genetic makeup of each sample. However, because strong evidence supports a history of multiple invasions of armigera , we alsober of K . We usedPP 1.1.2  and visuPP 1.1.2 .adegenet and ade4, as well as pairwise FST analysis [The number of clusters and the level of hybridization were also investigated using non-model-based methods such as Principal Components Analysis (PCA) with the R package analysis , 57. Addanalysis . PopulatH. armigera and H. zea in Brazil, we considered two groups of environmental variables in our analysis: climatic and land-use variables. For the climatic variables, we used elevation and 19 locality-specific bioclimatic variables from the WorldClim database, with a resolution of 30 arc-seconds [Given that populations of the two species are now in sympatry, interbreeding may occur at different rates, possibly related to the presences of their main agricultural hosts of soybean, cotton, and maize. To investigate the ecological context of hybridization of -seconds . To accoInstituto Nacional de Pesquisas Espaciais (INPE) and the United States Geological Survey – Earth Explorer (USGS/Earth Explorer), which provide images from CBERS 4 and LANDSAT 8, respectively.Land-use (i.e. landscape) variables were obtained classifying agricultural-landscape components such as soybean, maize, and cotton. Landscapes also contained other crops such as sugarcane, tree plantations, and orange orchards, as well as non-crop elements such as native forests, pastures, water, and urban sites that were also included in the classification maps. We quantified and characterized landscape attributes, using satellite images with a maximum of 2 months of differences in the collected data. This time window was necessary due to image quality, cloudy weather, and the availability of satellite images in public databases. Two databases were used for the satellite image collections, the We manipulated and classified the different attributes from satellite images using ArcGIS 10.2.2. Briefly, a buffer with a 25-km radius from the collection point was created to delimit the study area. This radius was chosen based on the relative size of regional agricultural areas, in order to prevent overlap between landscapes, and also based on the insect’s flight capacity (up to 1000 km) . Differe‘lmer’ function in the R package ‘lme4’ to estimate the relative importance of environmental factors for H. armigera and H. zea hybridization in Brazilian croplands [H. armigera. We tested the effect of landscape and climate, using these variables as fixed factors while controlling for the effect of the population (random = populations). We checked the significance of the models by evaluating χ2 and p-values from the likelihood-ratio test of model comparisons. The most complex models included the interaction between landscape and climate, followed by models of isolated factors, and naïve models.We constructed linear mixed-effects models using the roplands . The ave


In 2019, an outbreak of avian influenza (H3N1) virus infection occurred among commercial poultry in Belgium. Full-genome phylogenetic analysis indicated a wild bird origin rather than recent circulation among poultry. Although classified as a nonnotifiable avian influenza virus, it was associated with reproductive tropism and substantial mortality in the field. Of the 2 existing pathotypes, low pathogenicity avian influenza (LPAI) and highly pathogenic avian influenza (HPAI), infection with LPAI virus is mostly undetected in a flock but can cause some respiratory signs, egg drop, lethargy, and limited mortality , H3N1 virus was again detected on this farm –, panel AThe clinical picture in the field started mostly with discolored eggs for laying hens and breeders, followed by an increased mortality rate of up to 60% in breeder hens and 40% in laying hens. In addition, a marked egg drop, up to 100%, was reported. Recovered animals never fully regained their laying potential. A second wave of mortality caused by secondary infection was detected later. During autopsy, specific lesions were detected .,For the virus strain isolated in April from poultry on the index farm , hereafter called the 3497_April isolate, the IVPI was 0.13 , indicating a low pathogenicity phenotype. The complete coding sequences of the HA and neuraminidase (NA) genes, identified by Sanger sequencing, of the strains isolated in January  and April  confirmed their relationship. These results demonstrated that the virus from January 2019 had evolved into a poultry-adapted strain by April 2019, with a deletion in the stalk of the NA gene, previously described as crucial for adaptation from wild birds to poultry ,. Using n6 50% egg infectious dose (EID50) of the 3497_April virus, but no clinical signs were noted over 14 days. At the end of the experiment, 100%  and 90% (by standard hemagglutination inhibition [HI]) of the infected birds had seroconverted, confirming infection. Subsequently, to evaluate a possible synergistic effect of a potential bacterial cofactor, we inoculated a non–antibiotic treated lung/trachea homogenate from an H3N1 virus–confirmed animal simultaneously with the H3N1 isolate. At 6–11 days postinoculation (dpi), 40% of the birds exhibited clinical signs  and some died (10% on 13 dpi). This co-infection did not reproduce the clinical picture observed in the field, indicating that a bacterial cofactor would not be necessary to produce the clinical signs seen in the field.We performed in vivo infection studies  with the 3497_April isolate, obtained after inoculation of a lung/trachea homogenate into 9-day-old embryonated specific pathogen–free chicken eggs. First, we inoculated 11-week-old specific pathogen–free White Leghorn chickens oculonasally with 106 EID50/bird and 6 noninfected sentinel birds were added at 1 dpi. Of the 6 infected birds, 2 died at 6 and 7 dpi, showing either no or mild clinical signs at 5 dpi. Autopsy of each bird that died confirmed some of the lesions described for birds in the field . Among the sentinel birds, 1 bird died at 8 dpi without prior clinical signs or clear pathologic lesions during autopsy but demonstrated a higher amount of viral RNA (105) in the organs compared with the other sentinel birds at the end of the study (21 dpi)  , panel B<0.05) from 2 dpi to 5 dpi in both groups (nonparametric Wilcoxon signed-rank test). Excretion did not differ between sentinel and infected chicken except at 5 dpi, when cloacal excretion by sentinel chickens was significantly higher (p<0.05) than that by infected chickens (nonparametric Mann-Whitney test). No significant difference was observed between oropharyngeal and cloacal excretions, except at 5 dpi oropharyngeal excretion by sentinel chickens was higher (p<0.05) (nonparametric Wilcoxon signed-rank test). Analysis of viral RNA excretion , panel A,The viral RNA excretion period of the H3N1 virus under investigation demonstrates an extended period of virus excretion compared with that generally expected for LPAI viruses. LPAI virus excretion mostly starts 1–2 days after infection and continues for 7–10 days, peaking on day 5, then quickly declining because of immunity onset ,, panel BNo virus transmission to humans was reported. All 20 analyzed conjunctival swab specimens from farmers without clear influenza-like symptoms on the infected farms were negative for H3N1 virus by the Belgian National Reference Center of Influenza . Although this virus was classified as LPAI according to official definitions using IVPI or molecular criteria, it was more pathogenic in laying hens than expected for a non-notifiable LPAI. Increased pathogenicity was shown by systemic distribution, clinical signs, and increased transmissibility in poultry  under field and experimental conditions.


Culex modestus mosquitoes (3/239 pools) and isolates are related to strains from Volgograd, Russia. A screen for West Nile virus IgG was conducted and ~5% of the local human population tested positive.West Nile virus is widespread in southern Russia, where the fever appears annually. Since Western Kazakhstan borders on southern Russia, we examined mosquitoes in this region for the presence of West Nile virus. Virus was detected in a small proportion of  Flaviviridae family. It is a member of the Japanese encephalitis virus (JE) virus group, whose members include Japanese encephalitis virus, Saint Louis encephalitis virus (SLE) and Murray Valley encephalitis virus (MVE) (West Nile Virus (WNV) is a virus of arthropods that is classified within the us (MVE) . WNV is us (MVE) –5. In 19us (MVE) , 6–8. WNus (MVE) .objectives of our study were to determine: (1) whether WNV circulated in west Kazakhstan and (2) if so, what were the dominant species of mosquito involved in the region. We employed RT-PCR for viral RNA, followed by partial sequencing and phylogenetic analyses of isolates to determine how Kazakh strains may be related to strains from Russia and other parts of the world.West Nile fever has been diagnosed in 16 regions of the Russian Federation , severalMosquitoes for the study were collected using human landing catch approaches on collecting scientists, usually with a Mondchadskiy Bell . Collections took place on 28 different occasions in all 11 districts of WKO in the area between 49.0401 and 51.2520 degrees North and 47.3325 and 54.0851 degrees East between May 25 and July 09, 2015. This area is ~100 km south of Oral, Kazakhstan and ~600 km east of Volgograd, Russia. To detect viral RNA in mosquito pools, qRT-PCR analysis using viral E (envelope) gene sequences was carried out with a hybridization-fluorescent detection method, using the “AmpliSens WNV-FL variant FRT” kit (Russia). A Rotogene 6000 cycler was employed. This is a “single-tube” method, whereby RNA to cDNA to PCR product is carried out all in the same reaction mix.Two samples showing Ct scores of 26–28 were chosen for sequence analysis. The third positive sample had a Ct score of 36 and was not used.http://www.mbio.ncsu.edu/bioedit/bioedit.html) using 70 published genome sequences of WNV. Targeting conservative areas of the viral E gene with minimal nucleotide polymorphism, we selected four degenerate primers as follows:For sequence analysis, the above two samples were then subjected to further RT-PCR to generate a ~900b fragment from the E gene prior to sequencing. We designed primers for nested PCR with the BioEdit Sequencing Alignment Editor , as follows:Master Mix:Run Parameters:http://www.ncbi.nlm.nih.gov/) were used as comparison. Dendrograms were constructed using Mega 6.0 software, alignment of nucleotide sequences was carried out using the Muscle algorithm and phylogenetic trees were constructed using Maximum Likelihood  according to the manufacturer's instructions. Sequencing reactions were performed on an automated DNA Analyzer 3730xl (Applied Biosystems). Nucleotide sequences of about 800 bases were obtained and the sequences were placed in GeneBank . Sequences obtained in two rounds of PCR were combined into a common sequence using SeqScape 2.6.0 software (AppliedBiosystems). Reference strain sequences deposited in the international NCBI database   Branch swap filter—very strongSystem resource usage—1st+2nd+3rd + non-codingNo. of seqs—53No. of sites—692No. of bootstrap reps—1,000Following the mosquito findings, we ran a screen of 454 sera from inhabitants in the WKO, selected randomly. These serum samples had been collected by the Kazakh health authority in west Kazakhstan as part of routine screening activities and were kindly made available to us as anonymous numbered material . A commercial kit was used , following the manufacturer's instructions. These kits contain positive and negative controls and have the CE seal of approval from the EU. Briefly, 1:10 and 1:100 dilutions of sera were used in 96 well plates, in duplicate, and the assays were carried out twice. Final readings were the average of these four readouts.Mosquitoes were collected from 11 districts of the West Kazakhstan Oblast (WKO) and were identified by experienced entomologists, using Kazakh local morphology criteria. A total of 9,500 mosquitoes was combined into 239 pools. We identified insects in three genera and seven species. We did not differentiate between males and females.Culex, representing 56.2% of the total collection, and all in one species—Culex modestus. The composition of mosquitoes of the genus Aedes was more diverse and was represented by four species—Aedes flavescens, Aedes caspius, Aedes vexans, and Aedes cinereus. Mosquitoes of the genus Anopheles were less common in the region, with 1.8% of the total number of captured insects, represented by two species—Anopheles maculipennis (we did not differentiate between sl and ss) and Anopheles hyrcanus. In this study, only Culex modestus harbored WNV. Real-time analysis by RT-PCR revealed 3 positive samples for WNV RNA in Culex modestus found in the Akzhaiksky area  we were given were positive. Thus, while there have been no formal reports of WN fever in WKO, as stated earlier, it seems that a) the virus circulates in WKO and b) that people are being infected by it.C. modestus mosquitoes, but not in any others in our sample of positive insects.The main findings in this paper are that WNV circulates in NW Kazakhstan and is associated with This mosquito is a known WNV vector, is prevalent in south and southeastern Europe and seems to be moving North, perhaps in response to climate change, as mentioned earlier.The WNV strains we identified are very similar to those in Volgograd, Russia, about 600 km away from the collecting region in Kazakhstan.C. modestus mosquitoes. As a comparison, analogous reports of WNV prevalence in mosquitoes during outbreaks of disease in New York and Louisiana, USA, revealed mosquito infection rates similar to those found in this study  and there is limited travel between it and NW Kazakhstan. It seems more likely that people are being infected by resident is study , 18.It would have been useful for us to have been able to further test the individuals who were positive for WNV IgG, perhaps by looking for virus in blood, or in urine where the virus seems to persist for considerable times in certain cases . This hoCulex modestus in the northwest of Kazakhstan points to a focus of infection in this region. The public health authorities in the Republic will now hopefully mount careful monitoring of the mosquito population (particularly C. modestus) in WKO as well screening the population for signs of West Nile fever. Several other areas of Kazakhstan border with areas of Russia with known WNV cases, many relatively unpopulated, but some with cities  where continued exploratory work on WNV in Kazakhstan would be an important future public health activity.Detection of WNV in The datasets presented in this study can be found in online repositories. The names of the repository/repositories and accession number(s) can be found in the article/The studies involving human participants were reviewed and approved by Republic of Kazakhstan. Written informed consent for participation was not required for this study in accordance with the national legislation and the institutional requirements.All authors listed have made a substantial, direct and intellectual contribution to the work, and approved it for publication.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.


Picornavirales, while two are divergent and fall into the families Alvernaviridae and Reoviridae. However, a very recent study has suggested that there is extraordinary diversity in aquatic RNA viromes, describing thousands of viruses, many of which likely use protist hosts. Thus, RNA viruses are expected to play a major ecological role for marine unicellular eukaryotic hosts. In this review, we describe in detail what has to date been discovered concerning viruses with RNA genomes that infect aquatic unicellular eukaryotes.Increasing sequence information indicates that RNA viruses constitute a major fraction of marine virus assemblages. However, only 12 RNA virus species have been described, infecting known host species of marine single-celled eukaryotes. Eight of these use diatoms as hosts, while four are resident in dinoflagellate, raphidophyte, thraustochytrid, or prasinophyte species. Most of these belong to the order  Simple unicellular eukaryotes (protists) are supposed to have evolved over one billion years ago in the Proterozoic oceans ,2. UniceMarine unicellular protists include a heterogeneous collection of phototrophic  and heterotrophic  organisms with a wide variety of cell sizes and belonging to almost all eukaryotic lineages ,6. MarinIn the early 1980s, the new circumstances compelled a change, and many large and small DNA viruses were discovered with double-stranded genomes, ones that infect a cultivable, unicellular, eukaryotic-like variety of algal species such as a subset of green algae ,11. For Heterosigma akashiwo was isolated in 2003; it was reported as an example of an RNA virus infecting a marine protist . Hc. HcAlveremovirus . HcRNAV0viridae) . Two strviridae) .Reoviridae is a large family of viruses with genomes containing 10, 11, or 12 segments of double-stranded RNA (dsRNA). Micromonas pusilla reovirus (MpRV) is the first ds RNA virus yet discovered in the family Reoviridae that infects a protist  (The phyceae)  25]. Vi. ViReoviViral replication involves several steps including attachment to the host cell, penetration, uncoating, replication, assembly, and finally release . To dateHeterocapsa circularisquama . Th. ThNitzs viruses ,62. The Gonyostomum, Merotricha, and Vacuolaria are three representatives for freshwater; seven other genera: Chattonella, Chlorinimonas, Fibrocapsa, Haramonas, Heterosigma, Psammamonas, and Viridilobus have brackish species [Chattonella spp., Fibrocapsa japonica, and Heterosigma akashiwo. To understand the mechanisms of bloom formation, it is important to collect information on the raphidophyte life cycle, on cyst formation, and on vertical migratory behavior [Heterosigma infect the harmful bloom-forming raphidophyte, Heterosigma akashiwo (a member of the family Raphidophyceae), a marine alga with a world-wide distribution. Heterosigma akashiwo is a harmful bloom-forming alga living in temperate coastal waters. Since factors affecting its occurrence and bloom formation have remained elusive, the bloom’s timing and the severity of this eukaryotic algae are both still unknown [The Raphidophyceae are a small group of flagellated protists that inhabit diverse aquatic habitats . To date species . The Rap species . Some ofbehavior . Viruses unknown .Marnavirus. HaRNAV infects several strains of Heterosigma akashiwo from coastal British Columbian waters (Heterosigma akashiwo RNA virus (HaRNAV) belongs to the genus n waters . This wan waters . The genHeterosigma akashiwo isolated from Japanese waters, the northeast Pacific, and the northwest Atlantic [Heterosigma akashiwo viral pathogens and more importantly, sheds light on algal–virus pathogens and the complexity of virus–host interactions in the environmental protist [Marnavirus, to date [The infectivity of HaRNAV was tested against 15 strains of Atlantic . HaRNAV  protist . HaRNAV  to date .Thraustochytrium, in particular, have become of increasing interest to biotechnology research [Species of the genus research . The Thrresearch . The Labresearch .Schizochytrium had been previously accepted in the class Labyrinthulomycetes, family Thraustochytriaceae, within the kingdom Chromista. It has been reconsidered with a more precise classification of the Schizochytrium by supplementing its morphological, physiological, and molecular phylogenetic characteristic data [Schizochytrium sp. NIBH N1-27 [Aurantiochytrium, thereby establishing a new species for this virus. This virus received the name Aurantiochytrium single-stranded RNA virus 01(AuRNAV01).The genus tic data . SssRNAVBH N1-27 ,75. As a3‒6.4×104 infectious units per host cell [Marnaviridae [AuRNAV01, the first RNA virus infecting marine fungoid protists, was isolated from the coastal waters of Kobe Harbor, Japan, in July 2000 . All AuRost cell . The putaviridae ,75. Howeaviridae ,75.Dinoflagellates are a unique group of unicellular organisms classified among the harmful phytoplankton species. They are well known because of their having high morphological biodiversity and species richness, and several adaptation strategies to survive in various ecological niches. . They arHeterocapsa circularisquama Horiguchi is a harmful bloom-forming dinoflagellate that specifically infects and kills bivalves; there exist multiple viruses known to infect Heterocapsa circularisquama [Alvernaviridae and genus Dinornavirus. HcRNAV, a ssRNA virus specifically infecting Heterocapsa circularisquama, which was maintained in culture and isolated from the coastal waters of Japan. HcRNAV strains were grouped into two types, depending upon intra-species host-range analysis. These two types indicated complementary strain-specific infectivity. Typical strains of each type (HcRNAV34 and HcRNAV109) have been characterized. Both virus strains have capsids with icosahedral symmetry and a size of 30 nm in diameter, and their genome is ssRNA which is approximately 4.4 kb in size  has been identified as a major component of the picoplanktonic community in several oceanic and coastal regions [Prasinophytes (Chlorophyta) constitute a group of unicellular algae at the base of the green algal lineage. They include several marine photosynthetic picoeukaryote species described from cultured isolates ,124,125. regions .Micromonas pusilla reovirus (MpRV) is the first double-stranded RNA virus in the family Reoviridae and subfamily Sedoreovirinae genus Mimoreovirus that infects the photosynthetic marine picoflagellate M. pusilla [Reoviridae with 11-segmented genomes, namely the rotaviruses and aquareoviruses. It is noteworthy that the segment 1 of MpRV with 5792 bp has the longest open reading frame of any of the thus-far characterized reoviruses [Reoviridae with 11-segmented dsRNA virus within the enzyme core region of RdRp. The terminal sequences of MpRV are distinct from all those of sequenced viruses of the family Reoviridae. Phylogenetic analysis based on the RdRp sequences showed that MpRV cannot be grouped with any of the previously characterized genera of viruses and takes its position within another phylogenetic group. Therefore, MpRV was introduced as a member of a new and distinct genus designated Mimoreovirus [Micromonas pusilla RNA virus (MpRNAV-01B) or  pusilla . An 11 soviruses . MpRNAV reovirus . MpRNAV-reovirus .Marine microorganisms comprise a major portion of the living biomass on the globe, thereby driving ecological cycling and the flows of energy. While only comprising a small part of the total marine biomass, viruses dominate in abundance and genetic variability ,131,132.Screening each of the isolated viruses against other potential hosts than the original can answer important questions regarding protist viruses. No definitive relationship is yet known between diatoms and their viruses in situ; however, some authors believe that their ecological interactions are observable in nature, based on field surveys and physiological studies ,76. It iThe virus community can change the abundance and habitant of its hosts, meaning that diatoms have potential importance in controlling the quantity (biomass) and quality  of diatom populations in the natural environment . InnovatMarnaviridae (Picornavirales should reveal the secrets of their success. Although, Marnaviridae is definitely a major group, it only represents ~15% of the new marine viruses discovered in the recent metagenomic study [It is striking that such a wide variety of evolutionarily distant protist hosts are infected by the relatively closely related viruses of the family aviridae , and thaic study , many ofic study ,134.