Beyond the Pleasure Principle, which the founder of psychoanalysis wrote in the shadow of the virus. In reading Freud’s attempt to write a psychology of death in the context of this funereal period of history, I argue that he set out first, a mythological theory of viral law concerned with the death drive, before turning to second, a techno-scientific, biological theory of the same  law characterised by microbial immortality. Beyond this exploration of Beyond the Pleasure Principle, in the third part of the article I turn to a reading of Lacan’s interpretation of Freud’s work, where viral law becomes a story of cybernetics and nihilistic mechanisation. Here, perfect mechanisation, and the endless oscillation between message and noise, looks a lot like living death. Finally, I take up Derrida’s critique of Jacob’s molecular biology and, by extension, Freud’s theory of microbial immorality, that he thinks privileges an idea of repetitive sameness and opens up a space for cultural politics concerned with immunity against otherness. Derrida’s key point here is that this biological fantasy ignores the reality of viral sex that enables evolution to happen. What this means is that the other, even in its microbial form, is ever present, and that we must recognise the importance of difference to the possibility of social, political, and economic change.What is viral law? In order to being my discussion, I note that the last two years have been extremely difficult to understand and that we, meaning those who have lived through the pandemic, have struggled to make sense. Thus, I make the argument that the virus has impacted upon not only the individual’s ability to make sense in a world where every day routines have been upended, but also social and political structures that similarly rely on repetition to continue to function. According to this thesis, Covid-19 is more than simply a biological organism, but also a cultural virus that undermines the organisation of social, political, and economic systems and requires new ways of thinking about how we might move forward into a post-Covid world. In the name of beginning this project of making sense of Covid-19, I track back in history to the comparable reference point of the Spanish flu pandemic of 1918–1920 and, in particular, a reading of Freud’s  Contagious [seem like the best option?Looking back from the perspective of January 2022 it is uniquely difficult to make sense of the events of the last two years. We, meaning those who have lived through the pandemic, have been confused, disorientated, and perhaps even traumatised in subtle ways that mean that we can no longer make sense. We are desperately in need of what Priscilla Wald  calls anntagious  Wald traThis is precisely what the philosopher of illness Havi Carel  exploresNo Exit, Hell is simultaneously the other people one cannot escape, but also the lack of other people similarly confined. Hell is also endless monotony, the law of the same that produces indifference, boredom, and eventually the looming anxiety that emerges when one realises that the everyday life of banal routine that had kept the world turning no longer really exists. Under these conditions, even the medium-term future disappears from view. Thus, the future becomes a kind of black hole and the existential challenge becomes to shine a light into the void and begin to make sense. In Heideggerian and Sartrean terms, the challenge is for us to develop a project in order to move beyond what we might call ‘the viral end of history’.Moreover, this is not simply a problem for those struggling to breathe, or those suffering the debilitating effects of long Covid, who have seen the world disappear from view from the confines of their ‘sick rooms’, but also everybody else who has found themselves ‘locked down’, confined to home, and thrown back upon their own individual psychological reserves. In much the same way that we make sense together, developing a cultural world that enables us to understand events, we also endure in common, surviving the cruelties and turbulence of the world by supporting each other on an everyday basis. Basic routines shared in common enable us to maintain a sense that the world continues to turn and help us to understand the relationship between past, present, and future in order to construct meaningful lives. Although the phenomenologists who developed this theory of the ways in which we live in time, the existential tendencies of Heidegger and Sartre meant that they had little time for what they would have considered inauthentic or serial forms of temporality. However, under conditions of Covid lockdown even the most basic forms of everyday routine, which normally enable people to understand the link between past, present, and the future have collapsed to be replaced by a kind of grey monotony framed by the four walls of their homes turned carceral space. Caught in a situation resembling the scenario Sartre  sets up While this struggle for identity is one which everybody must face, we know that the post-pandemic future is not simply about the existential problems of the lonely individual. The precise reason I refer to the idea of ‘the viral end of history’ above is because it seems to me that the last two years might be best understood in terms of the definitive end of the end of history period first announced by Frances Fukuyama  in the lNow, in what follows I propose to construct a cultural politics of the Covid-19 pandemic in the name of opening up a debate about how we might move beyond the crisis into the future. In order to develop this cultural theory, I propose to track back in history to the last comparable reference point for making sense of Covid-19, the Spanish flu pandemic of 1918–1920. From this point, I move forward in history, offering a microbial, molecular, bio-political history of the 20th century moving into the 21st century. Following a discussion of the immediate post-World War I years marked by the Spanish flu, I consider the post-World War II period characterised by the emergence of molecular biology, the discovery of the DNA program, and what we might call viral sex. In the context of exploring this historical period, and in the name of setting out a cultural political theory of Covid-19 concerned with how we might think about moving beyond the pandemic, I base my discussion upon a number of key texts. As my discussion evolves these texts come together to form a coherent debate about the meaning of what I am calling ‘viral law’, which concerns thinking about the way the virus and viral society might move into the future on the basis of a law-like regulatory system.Beyond the Pleasure Principle in order to develop an initial theory of the meaning of viral law. My objective here is to set out a theory concerned with what Derrida [Beyond the Pleasure Principle is overwhelmingly coloured by his own feelings about death and the pervasive atmosphere of death, decay, and decrepitude that marked the period in which he was writing. My thesis is that in the face of this funereal atmosphere characterised by World War I and the death of his daughter Sophie, Freud ended up imagining the possibility of a completely immune ego, free from the horrors of finitude, in the form of a microbe, the famous protozoan discussed in chapter six of the text, which lives forever through the process of endless repetition and cellular division. Although Beyond the Pleasure Principle [Beyond the Pleasure Principle [Starting with the Spanish flu pandemic that raged across Europe and rest of the world between 1918 and 1920, and which Laura Spinney writes about in terms of a kind of void of immediate sense and historical memory, I read Freud’s  classic  Derrida  calls Frrinciple  is, in mrinciple  is markeBeyond the Pleasure Principle [Civilization and its Discontents [th April, 1945. It is in this context that I argue that Derrida sought to deconstruct Jacob’s similarly closed system of molecular biological programmatic life on the basis that what this theory really set out was an update on Weismann’s vision of the germ cell that lives forever, which Freud had adopted in his psychological defence against the funereal atmosphere caused by the Spanish flu pandemic. This is precisely why Derrida [theoretical possibility of total immunity against the outside. In this respect, the Freudian vision of somehow living beyond death, the Nazi fantasy of a Germanic utopia free from others, and Jacob’s molecular biological program that simply repeats ad nauseam, are impossible utopias that ignore the irreducible reality of communication, sex, and as a result death and finitude.Building upon my discussion of Freud’s rinciple , in partrinciple  critiquecontents , that we Derrida  shifts fBeyond the Pleasure Principle [Civilization and its Discontents [Beyond the Pleasure Principle [In order to begin my cultural political history of modern understandings of viral infection and pandemics, in this section of my article I return to the case of the Spanish flu of 1918–1920 and take up Freud’s rinciple , which wcontents , where Frinciple , my thesBeyond the Pleasure Principle [Viral Modernism [Writing on the relationship between Freudian psychoanalysis and the current pandemic, Brett Kahr  supportsrinciple  by paintrinciple  book, anrinciple , this quodernism , which wThe Second Coming [Before the invention of the electron microscope in the 1930s, it was, of course, impossible to observe the virus, meaning that it took on a kind of spectral, supernatural quality, that Woolf and the other modernists sought to represent through their own strange, delirious, texts. Unlike the contemporary Covid-19 period, where molecular biologists were able to observe, know, and sequence the virus more or less immediately, and populations could learn about and experience the contagious organism through the mediasphere, the invisibility, unknowability, and unpredictability of the Spanish flu virus meant that it was not possible to easily explain what was happening through normal narrative form. Instead, Outka shows how the modernists sought to reflect the social, psychological, and cultural impacts of the virus in the very ways in which they sought to make sense. Thus Woolf, Eliot, and Yeats would try to capture the way the virus corrupted the individual and destroyed previously reliable social structures through texts marked by an internal, formal sense of fragmentation, delirium, disorder, and a spectral atmosphere full of ghostly hauntings. As Outka  suggestsd Coming , Yeats id Coming , who annBeyond the Pleasure Principle [first viral law was the thanatological law concerned with the drive towards death and (self-)destruction. The reason I emphasise the word ‘first’ in this context is because we can see that Freud’s next step is to shift his focus back from the mythological figure of Thanatos to science and August Weismann’s microbial protozoan that lives forever through repetition, replication, and cell division in order to neutralise the mythic viral drive to extinguish life. In this second viral law, which takes the form of the simple, single celled organism, there is no contagion because there is no other, no sexual intercourse, and no space for infection.My key point is that I think that we might understand Freud’s rinciple  through Repression [Civilization and its Discontents [first viral law, the law of death and destruction, rules over the behaviour of the human-animal.Thus, we might conclude that Freud  imaginespression  and Civicontents , where FBeyond the Pleasure Principle [Civilization and its Discontents [We recall that this is precisely how Freud  opens uprinciple , and Frecontents  when he Totem and Taboo [In the face of his fear of nature, and his fear of the death drive which we suspect he might have imagined was embodied by the unknowable virus and the cold mechanisation of modern warfare, it is perhaps peculiar that Freud ends up tracking back through his own evolutionary story, comprising nd Taboo  with itsBeyond the Pleasure Principle [In responding to his own question about Freud’s survival techniques, Kahr  evokes nrinciple . PerhapsBeyond the Pleasure Principle [Beyond the Pleasure Principle [The Logic of Life [In the previous part of this article I presented a discussion of what I am calling viral law centred upon (a) Freud’s possible representation of the Spanish flu through the notion of the death drive, which potentially mirrored the experience of living in a society wracked by the virus in respect of the way in which it represented a demonic force relentlessly pushing forward towards death, and (b) his attempt to counter this funereal vision of life on the way to death through the development of a second viral law based upon reference to August Weismann’s biology and particularly the figures of the germ cell and protozoan that reproduce endlessly without the need for otherness, sex, or death. My argument here was that ultimately Freud sought to immunise himself against the atmosphere of death circulating from 1914 through to 1920 and the publication of rinciple  and thatrinciple  cybernetrinciple  in his srinciple  seminar  of Life .Life as Surplus [Logic of Life [Beyond the Pleasure Principle [The Freudian Robot [As a result, the point of this section of the article is to project the Freudian theory of viral law set out in the second part of the piece into a reading of structuralist and post-structuralist thought concerned with the emergence of cybernetics and information theory, post-industrial high-tech capitalism, and the new field of molecular biology, which taken together would later lead to the emergence of neoliberal bio-capitalism Melinda Cooper explores in her book,  Surplus . Given t of Life , brings rinciple  to cyberan Robot  Lydia Lian Robot  reading Beyond the Pleasure Principle [However, before we reach Derrida’s critique of molecular biology and Freud’s viral law, I propose to turn towards Lacan’s  reading rinciple  and the The Invasion of the Body Snatchers, and we might add the original version of The Thing later remade for the bio-capital age by John Carpenter, Wald settles on William Burroughs as the key critic of the post-war American virus of conformity and control, citing works such as The Ticket that Exploded [simply a biological problem, but also an issue that impacted upon the social, political, economic, and cultural systems that were essentially concerned with the circulation of information. In the wake of the post-war cybernetic revolution and Watson and Crick’s discovery of DNA in 1953, this was, of course, the essential structuralist point. As it became increasingly clear that life itself was an information system that could be understood in terms of signs, symbols, and code, and on the basis of the idea that the social, political, economic, and cultural systems could similarly be understood in terms of information transfers and feedback systems, the structuralists began to develop a vision of a system capable of taking in understandings of the natural and human worlds drawn from a range of different academic disciplines, including semiotics, sociology, history, psychoanalysis, literature, and biology. We know that Lacan’s particular contribution to this field was to re-read psychoanalysis through cybernetic and information theory and seek to project the Freudian notion of the unconscious onto the language systems that enables the individual to operate in social context.We know that this American tendency to seek immunity and screen out otherness in the name of protecting some sense of essential identity was subject to critical scrutiny long before Derrida’s critique of Jacob’s techno-scientific, cybernetic vision of the living that sought to marginalise relationality. Referring to science fiction films, such as Exploded  and the Beyond the Pleasure Principle [Beyond the Pleasure Principle [In this regard, Lacan  starts hrinciple , Freud’srinciple  revisionrinciple  and the Beyond the Pleasure Principle [and a perfectly functional machine able to keep working endlessly into the future , throws up is the precise problem of human freedom somewhere between simple microbial life that lives in order to divide and divides in order to live with no need for otherness, sex, or death and the artificial techno-scientific mechanical simulation of life  that works in order to works and keep working and nothing more.Although there is no true sense of freedom in Lacan’s real, where the human is a body in pieces, his (Lacan’s) first machine, the imaginary mirror relation that alienates the subject from themselves, subordinates humanity to the image. While the subject might prefer to remain inside this basic circuit of the real and imaginary self, Lacan says that the unconscious dream machine ensures that they are constantly reminded of their lack caused by their alienation from nature (mother) and leap into language and symbolic civilization. This is when we (humanity) find ourselves caught up in the repetitious logic of drive and the virus that is precariously balanced on the edge of life and death, being and nothingness, and the relentless pursuit of some other that could make the self complete. As we have seen above, this pursuit of the other, in the form of an endless parade of signs, boils down to the basic mechanism Freud identified in rinciple . Everythrinciple  move, whOf Grammatology [Finnegans Wake [Memoirs of My Mental Illness [It is precisely this problem of freedom that Derrida  addressematology  is no loans Wake , which h Illness , which mBeyond the Pleasure Principle [Thus, the program represents a kind of unconscious, deep- or infra-structure that provides the basic instructions for the reproduction of life. Although change is possible through copying errors and selection, Jacob is clear that this system is primarily about repetition and that evolutionary change happens only very gradually through advantage gained in the wake of accidental mutation. Akin to Weismann’s germ cell, Freud’s concept of the death drive, Lacan’s cybernetic switching system, and what we have called viral law, Jacob’s program is not subject to conscious transformation or wilful changes of course. The key point here is that there is no freedom, no other, no outside in Jacob’s molecular biology. According to this thesis, Jacob repeats Weismann’s conclusion, which Freud would in turn repeat in chapter six of rinciple , that seBeyond the Pleasure Principle [Of course, Derrida’s  point isrinciple , Derridarinciple  basic prBeyond the Pleasure Principle. In identifying the same kind of deep structure operating in Jacob’s [It would seem to me that this is precisely what Freud  trades a Jacob’s  moleculaWhile this process looks like a hostile take-over from the point of view of the unicellular utopia of the same committed to programmatic reproduction without difference, Derrida’s point is that what Jacob admits through his recognition of molecular sex is the fact that difference, otherness, sexuality, and as a consequence the metabolic process of death, exist on the most basic level of the living. In other words, there is no sense in which primordial, molecular life is self-identical, programmatic and possessed by the drive to the endless reproduction of the same, whereas later, more evolved complex life is subject to laws of otherness, relationality, sexuality, and finitude. On the contrary, what Derrida  takes frand a funereal vicious circle comprising a monstrous confusion of life and death. Instead, Derrida’s virus is the primordial supplement that introduces sex, death, and the possibility of evolutionary transformation into what would otherwise look like a completely closed system of the living unable to develop and change. In this respect we might conclude that the idea of the virus, and the notion of viral law, speaks to Derrida’s concept of the pharmakon, which is simultaneously a cure and a poison. In this interpretation, which recalls Carl Zimmer’s [Beyond the Pleasure Principle [In Derrida’s  reading Zimmer’s  point abrinciple  in orderWe have seen that this situation may have led Freud to mythologise the flu virus in the idea of the death drive, only to then take off in an alternative direction in order to suggest a utopian story of unicellular immortality and unstoppable life that knows no other. Following Lacan’s re-reading of the death drive in terms of the idea of a perfect cybernetic system that seems to destroy the possibility of life in a kind of terminal feedback loop, we took up Derrida’s reading of Jacob’s molecular biology, which takes in Freud’s idea of microbial immortality and builds upon Lacan’s insight that the perfectly functioning machine ends up creating a state of living death. As a result, Derrida critiques Jacob’s molecular theory of programmatic life as a fable of the possibility of endless generation, growth, and development without the limit of otherness or sexuality, which necessarily introduces the brake of finitude and death. It is in responding to this bio-cybernetic fantasy, which, if we were to evoke Heidegger  we might


Second,L'Abate, , p. 14. L'Abate, . In factL'Abate, . Each stL'Abate, .vice versa. In this regard, Colesso et al. (Some studies have supported this theory (L'Abate and Wagner, o et al.  have diso et al. .transmission of particular relationship styles or symptoms from one generation to another, including the present situation in the family of procreation” (L'Abate, Moreover, according to L'Abate , relatioL'Abate, , p. 125.L'Abate, . Hence, L'Abate, . ExperieL'Abate, . On the L'Abate, . ExperieL'Abate, .The mechanisms underlying the intergenerational transmission of relational styles may be related to the fact that the experience of specific parental relational styles may lead children to learn that style as an appropriate strategy (Festinger and Carlsmith, even though they derive from different theoretical sources, predict seemingly similar patterns of relationships between partners or among intimates” (L'Abate et al., The RCT might be misunderstood as “an old wine in a new bottle” since it seems to present theoretical convergences with two other important theories, namely, attachment and social learning theories Bowlby, . In factIn contrast, RCT seems to present convergences and divergences with social learning theory Bandura, . In factA better comprehension of the intergenerational transmission of relational styles could lead to several clinical implications. First, identifying both the relational styles patients experienced in childhood and the ones they act out in adult relationships may contribute to acquire information regarding the type of psychological interventions. For example, the AA style seems to be the most dysfunctional one as well as the most difficult one to change L'Abate, ; thus, iProtective factors should be also targeted in the clinical practice. The self-importance of patients, which, according to RCT, corresponds to the importance a person attributes to the Self, represents a core factor in this regard L'Abate, . In factad hoc psychological interventions targeting the relational competence and styles of patients (L'Abate et al., These considerations could help clinicians to plan Overall, according to the intergenerational transmission of relational styles (Cusinato, Moreover, it is important to highlight that recent research stressed out to employ a bidirectional approach to intergenerational transmission according to which caregivers and children influence each other in a reciprocal way (De Mol et al., The intergenerational transmission could have implications on both the research and the clinical level. In fact, the RCT gives importance to intimate relationships with others, as central elements for the development of personality and relational styles; for this reason, it could be useful in couple and family therapies L'Abate, .Moreover, RCT allows a better understanding of the influence of relational competence and styles for the sense of Self of patients, significant relationships, and symptoms, thus, weighting them in psychological interventions. In addition, future studies should investigate the relationship between attachment and relational competence. In fact, if this relationship is proven, when knowing how individuals interact with others, it may be possible to have information on the attachment of individuals as well.In conclusion, future studies should deepen both the intergenerational transmission of relational competence and styles and all the protective factors that may buffer the transmission of dysfunctional relational styles across generations.FT and AAR conceived and wrote the manuscript. SM provided important intellectual contribution and critically revised the manuscript. All authors approved the final version.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.


Antimicrobial resistance (AMR) is a global public health problem affecting animal and human medicine. Poultry production is among the primary sources of income for many Zambians. However, the increased demand for poultry products has led to a subsequent increase in antimicrobial use. This study assessed the awareness of AMR and associated factors among layer poultry farmers in Zambia. A cross-sectional study was conducted among 77 participants from September 2020 to April 2021. Data was analysed using Stata version 16.1. The overall awareness of AMR among the farmers was 47% (n = 36). The usage of antibiotics in layer poultry production was high at 86% (n = 66). Most antibiotics were accessed from agrovets  and pharmacies  without prescriptions. Commercial farmers were more likely to be aware of AMR compared to medium-scale farmers , as were farmers who used prescriptions to access antibiotics compared to those who did not , and farmers who did not treat market-ready birds with antibiotics compared to those who did . The awareness of AMR among some layer farmers was low. Therefore, policies that promote the rational use of antibiotics need to be implemented together with heightened surveillance activities aimed at curbing AMR. The use of antimicrobials in layer poultry production has continued to increase significantly in the recent past as the demand for poultry meat and eggs increases due to improvements in the social and economic lives of people . AntimicEscherichia coli is one of the highly antimicrobial-resistant pathogens in livestock [Enterococcus and Salmonella have been reported in livestock production [Staphylococcus aureus and Listeria species have developed resistance to antimicrobials that are commonly used in poultry and humans [Humans can contract antimicrobial-resistant microorganisms from animals through the food chain ,15,16. Eivestock ,20,21,22oduction ,23,24,25d humans ,27,28. TMany poultry farmers have access to antibiotics without prescription ,31,32,33Since AMR has been shown to affect animals, humans and the environment, there is a need to address this problem using the “One Health Approach” . Under tAt a global level, a lack of awareness of AMR and associated factors among poultry farmers has been reported to be among the factors that exacerbate AMR . PoultryIn Zambia, poultry production is a source of income for many farmers and contributes to the country’s food security . There hThis study was conducted to assess the awareness of AMR and associated factors among layer poultry farmers in Zambia.Of the 77 layer farmers interviewed, the majority  were male. About 22 (28.6%) were from Kitwe, and 24 (31.2%) sourced antibiotics from agrovet shops. A total of 39 (50.7%) participants were commercial farmers , 66 (85.7%) used antibiotics, 39 (50.6%) used a prescription to access antibiotics, 45 (58.5%) used antibiotics for the prevention of infections, 66 (85.7%) consulted a veterinary doctor before using antibiotics, and 48 (62.3%) observed the antibiotic withdrawal period. Additionally, 55 (71.4%) farmers did not treat market-ready birds with antibiotics, and 70 (90.1%) practised biosecurity. There was evidence of an association between awareness of AMR and district of residence, type of farmer, source of antibiotics, use of a prescription to access antibiotics, consultation of a veterinary doctor before using antibiotics, knowledge of the observation period, treatment of market-ready birds and biosecurity practice. The overall awareness of AMR among study participants was 46.8% (n = 36), as shown in The results from a multivariable analysis of factors associated with awareness of AMR are shown in This study aimed to assess antimicrobial resistance (AMR) awareness and the associated factors among layer poultry farmers in Zambia. The overall awareness of AMR among the layer poultry farmers was 46.8%. Factors associated with awareness of AMR in our study included type of farmer , source of antibiotics , use of prescriptions to access antibiotics and avoiding the use of antibiotics to treat market-ready birds.Less than 50% of the participants in the current study were aware of AMR and the associated factors. These results corroborate the findings in similar studies conducted in low- and medium-income countries, where the majority of the participants were not aware of AMR and the associated factors ,42,49. ACommercial farmers from different districts were more aware of AMR than medium-scale and small-scale farmers. Similarly, a study in Ghana reported that commercial farmers tend to be more aware of AMR and antibiotic use than medium and small-scale farmers . In low-Our study found that farmers who accessed antibiotics from agrovets were more likely to be aware of AMR than those who only went to the veterinarian or general pharmacies. This could be because agrovets are more accessible than veterinarians and general pharmacies and there are more agrovets in many areas than veterinarians and general pharmacies . SimilarAntibiotics were used by 86% of the layer poultry farmers and were mainly accessed through agrovet shops and veterinarians without using a prescription. Similar findings have been observed in some studies conducted in other countries, including Ghana, Kenya and Grenada ,49,59. IOur study found that most layer poultry farmers consulted veterinarians on antibiotics in poultry. Despite accessing antibiotics from various sources, the participants reported that they consulted veterinarians on antibiotics used in poultry. Similarly, a study conducted in Ghana showed that many poultry farmers consulted veterinary officers on antibiotics . ConsultOur study revealed that many farmers implemented and practised suitable biosecurity measures on their farms. This is good for the layer poultry farmers because biosecurity measures help prevent the transmission of infections from humans to animals and vice-versa. The biosecurity measures included the fencing of poultry, footbaths at the gate, restriction on poultry entrance, limited access to poultry by other animals and isolation of sick birds. Biosecurity measures in poultry farming are crucial for disease prevention in poultry and a consequent reduction in the use of antimicrobials ,66. HoweThis study had some limitations that must be considered when interpreting our findings. The study used a small sample size of the poultry farmers that were registered with the animal health authorities of Lusaka and Copperbelt provinces at the time of the survey. However, to the best of our knowledge, this study was the first to be conducted in Zambia to pave the way for the development and implementation of AMR surveillance strategies in layer poultry farming. Thus, this epidemiological survey will be combined with molecular methods that will help come up with the best ways of monitoring AMR in layer poultry production in Zambia.A cross-sectional study was conducted in Zambia’s Lusaka and Copperbelt provinces from September 2020 to April 2021. Lusaka, Kafue, Rufunsa, Chongwe, Kitwe, and Ndola cities were purposively selected from the two provinces after considering the similarities in farming activities, practices, and population density based on the Poultry Association of Zambia (PAZ) data for layer poultry farms . The mapThe study was conducted among eligible layer poultry farmers in the study sites. To be eligible, a farmer had to reside in Lusaka or Copperbelt provinces and sign a written consent to be part of the study. All the farmers reared layer chickens in the production stage at the time of data collection. We excluded layer farmers who were not available during the study period and those who were not comfortable being interviewed due to the fear of contracting COVID-19. We also excluded layer farmers who reared layer chickens that were not in the production stage.A multi-stage sampling procedure was used in this study. The districts in Lusaka and Copperbelt provinces were categorised based on farming activities and practices. Lusaka province had seven (7) districts, while the Copperbelt had 10 districts. Then, we purposively selected a total of six (6) districts from the two (2) provinces. In Lusaka province, the selected districts included Chongwe, Kafue, Lusaka and Rufunsa whereas the selected districts from the Copperbelt province were Kitwe and Ndola. Research assistants were first assigned in each province to identify potential participants from the eligible farms in each selected district. Registers from PAZ and District Veterinary Offices (DVOs) revealed a total of 96  layer poultry farms. In each of the selected districts, farms were categorised into three (3) strata, i.e., commercial farms , medium-scale farms  and small-scale farms (≤1000 birds). Of the 96 farmers that were identified, 92 met the inclusion criteria. Since the obtained number of layer poultry farms was small, we conducted a complete enumeration. Therefore, we aimed to enrol all the farmers that were identified through the registers and met the inclusion criteria. Overall, 77 eligible layer farmers were included in the study and completed the questionnaire.The data were collected using a semi-structured questionnaire adapted from a study by Nkansa and colleagues . Firstly® and imported into Stata® version 16.1 . Categorical variables were expressed as frequencies and percentages. The test of associations was done using the Pearson chi-square test and, where necessary, Fisher’s exact value.For statistical analyses, the collected data were entered into Microsoft Excelp < 0.20 from the univariable analysis to obtain adjusted odds ratios. The multivariable regression model was fitted using a machine-led backward stepwise regression technique. The final model was fitted using robust standard errors to account for clustering among the farmers from similar farming blocks. The Hosmer–Lemeshow goodness-of-fit test was used to assess the predictive ability of the model. Since the model fit was inadequate, we further investigated the possible interactions between significant variables and none were found to reach any statistical significance. Additionally, we assessed for multicollinearity using the variance inflated factor (VIF), and the highest value was 3.54, suggesting that multicollinearity was not a problem. All statistical tests were done at a 5% significance level and a 95% confidence level.For the study outcome (awareness of AMR), univariable logistic regression was performed with the study characteristics to obtain crude odds ratios. Further, a multivariable logistic regression model was fitted, including only variables with a The study found low awareness of AMR and associated factors among layer poultry farmers in Zambia. These findings indicate the need to provide education to the farmers on AMR and associated factors. There is a need to develop and implement AMR surveillance and antimicrobial stewardship programs in layer poultry production in Zambia.


BAP1, CDKN2A/B, and the TERT promoter, which are associated with particularly aggressive tumor biology. Despite the evident clinical value, advanced molecular profiling methods are not widely incorporated in routine clinical practice for meningiomas.Meningiomas are the most common primary central nervous system (CNS) tumor. They are most often benign, but a subset of these can behave aggressively. Current World Health Organization (WHO) guidelines classify meningiomas into three grades based on the histologic findings and presence or absence of brain invasion. These grades are intended to guide treatment, but meningiomas can behave inconsistently with regard to their assigned histopathological grade, influencing patient expectations and management. Advanced molecular profiling of meningiomas has led to the proposal of alternative molecular grading schemes that have shown superior predictive power. These include methylation patterns, copy number alterations, and mutually exclusive driver mutations affecting oncogenes, including To assess the degree of concordance between the molecular profile of meningiomas and the histopathologic WHO classification, the current method of predicting meningioma behavior.In a two-year single-institution experience, we used commercially available resources to determine molecular profiles of all resected meningiomas. Copy number aberrations and oncogenic driver mutations were identified and compared with the histopathologic grade.TERT, CDKN2A/B, or BAP1 of which 6% occurred in grade 1 meningiomas.One hundred fifty-one total meningioma cases were included for analysis . Chromosomal analysis of 124 of these samples showed that 29% of WHO grade 1 tumor featured copy number profiles consistent with higher grade meningioma, and 25% of WHO grade 2 meningiomas had copy number profiles consistent with less aggressive tumors. Furthermore, 8% harbored mutations in Routine advanced molecular profiling of all resected meningiomas using commercially available resources allowed for identification of a significant number of meningiomas whose molecular profiles were inconsistent with WHO grade. Our work shows the clinical value of integrating routine molecular profiling with histopathologic grading to guide clinical decision making. Meningiomas, named for their cell of origin, are the most common intracranial central nervous system (CNS) tumors in adults representing approximately one-third of all primary adult CNS tumors –3. The WIn current practice, WHO grade and extent of resection are the most widely used metrics to predict tumor behavior and guide management of meningiomas , 7. MostA developing understanding of the molecular landscape of meningioma suggests that WHO grade alone may not provide an adequate prediction of tumor behavior, for surveillance or adjuvant treatment planning considerations. Several recent publications have proposed alternative grading systems that incorporate genomic and/or epigenomic data in order to better predict meningioma behavior, particularly targeting those that do not behave in concordance with their assigned WHO grade –14. ThesBAP1, CDKN2A/B, and mutations within the promotor region of the TERT genes (via vendor) enhancing their appeal  of the University of Oklahoma Health Sciences Center (OUHSC) (IRB protocol number 10195). All patients who underwent their index surgery with a histopathologic diagnosis of meningioma following CNS tumor resection from May 2019-April 2021 were included in the study. Patient data including demographics, tumor location, and history of prior resection were collected by retrospective chart review.th edition of the WHO Classification of Tumours of the CNS published in 2016 , posterior convexity , falcine/parasagittal, tentorial, tentorial sinus, peritorcular, intraventricular, or multifocal. Meningiomas were further classified based on their sublocation  . Summary statistics are reported as counts or proportions for categorical variables. For continuous variables  mean and standard deviation are shown in figures. Statistical analysis of copy number alterations and Ki-67 between meningiomas with high-grade and low-grade copy number variations were done by one-way analysis of variance (ANOVA) with Tukey’s NF2 gene, the most common genetic disturbance associated with meningiomas, were evenly distributed across grades and comprised approximately 40% of the total meningioma cohort. The common histopathologic subtypes were all represented in our cohort with meningothelial and transitional comprising the majority of grade 1 tumors  occurring in WHO grade 1 meningiomas  with an altered TRAF7 or AKT1 E17K with an altered TRAF7 . This is consistent with a recent report that applied advanced genomics to create a molecular grade with improved prediction of meningioma behaviour, with up to 32% of cases reclassified when molecular data were applied . There aIn recent years, the clinical impact of molecular data in the treatment and management of meningiomas has become evident. Numerous classifications that incorporate molecular data have been shown to improve the prediction of tumour behaviour for meningiomas. With the rapid advances in molecular understanding of meningiomas over the recent years, it will be important to determine how this information can be integrated in routine clinical settings and standardized nationally, in particular for institutions with limited resources, and to correlate this information with clinical outcomes.The original contributions presented in the study are included in the article/The studies involving human participants were reviewed and approved by Institutional Review Board University of Oklahoma Health Sciences Center. Written informed consent for participation was not required for this study in accordance with the national legislation and the institutional requirements.Study design and data analysis: AR, PP, and ID. Histopathologic grading, processing, and shipment of meningioma samples for advanced molecular profiling: JP and K-MF. Drafting of manuscript: AR and PP. Proofreading, finalizing, and approval of manuscript: all authors.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.


A critical strainrate of the electrospinning jet is determined by theoretical modelingat which point the polymer network undergoes a stretch transitionin the fiber direction, resulting in a high molecular orientationthat is partially retained after solidification. Nearing a nanowireboundary, local adsorption of the polymer and hydrodynamic drag furtherenhance the molecular orientation. These theoretical predictions aresupported by polarized scanning near-field optical microscopy experiments,where the dichroic ratio of the light transmitted by the fiber providesevidence of increased orientation nearby nanowires. The addition ofnanowires to enhance molecular alignment in polymer fibers might consequentlyenhance properties such as photoluminescence quantum yield, polarizedemission, and tailored energy migration, exploitable in light-emittingphotonic and optoelectronic devices and for sensing applications.Themolecular orientation in polymer fibers is investigated forthe purpose of enhancing their optical properties through nanoscalecontrol by nanowires mixed in electrospun solutions. A prototypicalsystem, consisting of a conjugated polymer blended with polyvinylpyrrolidone,mixed with WO For instance,nanofibers undergo substantial elongation during the electrospinningprocess, resulting in diameter shrinkage and preferred molecular orientationin the direction of the extensional flow.7 Rapidsolvent evaporation, accelerated by the high surface-to-volume ratioin such jets, leads to rapid nanofiber solidification that retainsthe oriented conformation. In the resulting fibers, the mechanical,electrical, and optical characteristics of polymers may so be enhancedcompared to the bulk.10 Importantly, further effects can be obtained by mixinghard fillers, such as nanowires (NWs), nanotubes, or nanoparticles,with the polymer solution.17 The question arises whether such fillers influence the polymer molecularorientation and properties in the resulting solid nanofibers, andif so, what are the physical mechanisms involved?Polymernanofibers are valuable building blocks for a variety ofapplications, ranging from smart textiles to electronics and photonics.9 thesepolymers are useful for light-emitting diodes,18 lasers,19 and field effect transistors.20 The molecular conformation of conjugated polymers,specifically the length, orientation, and density of their conjugatedchain segments, has a strong impact on their photophysical properties.26 For example, when the polymer chromophores (molecular parts thatgenerate color) are aligned along the axis of a nanofiber, the photoluminescencequantum yield and the degree of polarization of the emission are significantlyenhanced, leading to miniaturized light sources with improved signal-to-noiseratios of detected optical signals.27 Moreover,hybrid systems composed of organic semiconductors and of inorganicnanowires with complementary properties have been exploited in solarcells, light-emitting diodes, electrochromic devices, and photocatalyticsystems.29 Whenever adding NWs to electrospun solutionsleads to enhanced alignment of conjugated polymers, this can potentiallyimprove the ability to funnel light to chromophores oriented alongthe fiber length,30 a condition highlydesirable for light-harvesting applications,31 and for enhancing the polarization degree of the light emitted bythe hybrid nanofibers.The effectsof such local variations in molecular conformationin electrospun conjugated polymers are especially interesting. Giventhe conjugated bonding along their chain backbone,The interface between solid fillers anda solution entails adsorptionand hydrodynamic interactions, which affect the polymer molecularconformation. Generally, in the vicinity of an obstacle such as afiller, polymer chains in a solution are confined by the obstacleboundary in the directions normal to it. In terms of chain conformation,this means that the probability that the chain will expand in thatdirection is low. Consequently, the probability of chain expansionin the other directions is augmented. In the case of NW fillers, boththe chain extension and orientation might increase in the directionalong the NW. In addition, close to a filler boundary, the flow ofthe electrospun jet may be affected. The flow gradient (strain rate)that is responsible for chain extension can be locally decreased orincreased by the hydrodynamic friction (drag) exerted by obstacles,thereby impacting molecular orientation.33 These calculations showed that chains adsorbed by an attractingsurface crowd near the surface into a flat pancake shape, to compensatefor the loss of entropy due to confinement. Recent studies on polymerinteractions with nanoparticles, by small-angle neutron scatteringof polystyrene solutions loaded with silica nanoparticles,35 and by molecular dynamics simulation of polymer solution with nanorods,36 demonstrated extended conformation of chainsclose to a filler boundary. Electrospinning of poly(ε-caprolactone)solutions with multiwalled CNTs exhibited the nucleation and formationof chain-folded crystals on the CNTs, with distinct orientation.37 However, these studies do not provide simpleand practical rules for achieving desired or optimal molecular orientationfor optical applications by electrospinning. Therefore, it is ourintention to offer a theoretical analytical solution, based on a scalingapproach, which correlates the extensional flow conditions of electrospinningwith the molecular orientation induced by chain extension, and validateit by optical measurements.The interaction betweenpolymer chains and a confining surfaceand its effect on chain conformation have been a long standing topicin polymer physics since the seminal works of Flory and de Genneson polymer chain adsorption, using scaling and mean-field calculations.3) NWs. Blending conjugated polymers, which typically haverigid backbones and relatively low molecular weight and level of entanglement,with PVP, turns out to help obtain electrospun fibers without defectsor beads.38 Moreover, in light-emittingconjugated polymers, the intrachain and interchain coupling betweenchromophores can favor the energy transfer from the photoexcited chromophorestoward those with a lower energy, resulting in red-shifted emission.Such funneling of the excitation toward the chromophores with a lowerenergy is also expected to decrease the polarization degree of theemission.39 Therefore, the presence ofPVP may also be beneficial for improving the polarization degree oflight emission because of a decrease of interchain coupling in MEH-PPV.Here, we develop a theoretical modeland study the exemplary solutionof poly (MEH-PPV)and polyvinylpyrrolidone (PVP) added with tungsten trioxide , which provides maps of the optical dichroicratio of MEH-PPV, supporting the theoretical predictions.Chain extension is analyzedby calculating the stretching and retractingdynamics of chains and the different effect on each polymer speciesin the blend. Monomer alignment probabilities are used to assess theadsorbing effect of obstacle boundaries, and the hydrodynamic draginduced by the NWs is used to assess the local increase or decreaseof the strain rate. Effects are combined to calculate and map themolecular orientation at variable distances from NWs. Experimentalevidence is obtained from nanofibers electrospun from the PVP/MEH-PPV/WOTheresults demonstrate that the incorporation of nanowires innanofibers made of conjugated polymers turns out to be effective forenhancing the alignment of the polymer chains at the nanoscale, comparedto neat polymer nanofibers. As a result, these nanowire-in-nanofiberhybrids are highly suitable for investigating nonradiative energytransfer and charge separation effects in nanocomposite systems andfor exploitation as building blocks of miniaturized light-emittingdevices with polarized emission and light-harvesting devices. Thecomprehensive rationalization of the molecular alignment mechanismsin the nanocomposite nanofibers demonstrated here can establish newdesign rules for next light-emitting nanostructures realized by electrospinningtechnologies.22.1MW = 150–250kDa, Sigma Aldrich) and 30 mg of PVP  in 1 mL of a mixtureof chloroform  and dimethyl sulfoxide with a volumetric ratio of 9:1 (v/v). The solutionsare stirred for 24 h. Next, WO3 NWs (Sigma Aldrich) areadded to the solution to obtain a weight ratio of 20% with respectto the dry weight of the PVP/MEH-PPV mixture. The solutions are sonicatedfor 2 h and then injected through a 21G needle with a flow rate of2 mL/h by using a programmable syringe pump (Harvard Apparatus). A7 kV voltage bias  is appliedbetween the needle and a drum with a grounded disk (8 cm diameterand 1 cm thickness) rotating at 2000 rpm and positioned at a distanceof 5 cm from the needle. For optical characterization, fibers aredeposited on quartz substrates .Solutions are preparedby dissolving 30 mg of MEH-PPV  through a 20× objective . The fluorescence is collected by the same objectivein a backscattering configuration, and the intensity is measured witha photomultiplier. In addition, the intensity of the excitation lighttransmitted by the sample is measured in each scan with a second photomultiplier.The morphology of the composite fibers is investigated using anFEG-SEM  at acceleration voltages in the interval 1.5–10kV. Fluorescence confocal microscopy is carried out by using laserscanning (Olympus FV1000) coupled to an inverted microscope. Samplesare excited using an Ar2.39 The microscopeoperates in the emission mode, where a probe made of a tapered opticalfiber  illuminatesthe sample in the near field, while the signal transmitted by thesample is collected using an aspheric lens and measured with a miniaturizedphotomultiplier (Hamamatsu R-9880). Topography maps, h, representing the localthickness of the sample, are measured simultaneously with the opticalsignal in each scan by the shear-force method. For nanoscale opticalimaging, a solid-state laser , modulated in amplitude and polarization, is coupled to thetapered fiber. The polarization of the laser is controlled by meansof a photoelastic modulator , which operatesas a waveplate with a retardation modulated sinusoidally at a frequency, f = 50 kHz. As a result, the ratio between polarizationcomponents of the light coupled into the probe optical fiber, measuredalong two mutually orthogonal directions, acquires a 2f modulation. The amplitude of the laser is instead modulated at frequency f* (f*/f < 0.1) byusing a chopper (Scitec Instruments 300 CD). The photomultiplier isconnected to two different digital dual lock-in amplifiers (StanfordResearch SR830DSP): while the first one, which is referenced to f*, is used to obtain the optical transmission signal averagedover all polarization states (IDC), thesecond lock-in that is referenced to 2f providesan output signal (IAC) representativeof the sample response to polarized radiation. The extinction coefficientis then evaluated through the expression TNF = – ln [IDC]/h, whereas the dichroic ratioof the sample, I∥ and I⊥ are the transmitted intensitiesfor polarization aligned along two mutually orthogonal directions,is quantitatively evaluated from the ratio IAC/IDC. While this ratio accountsfor local variations of the intensity of nonpolarized scattered lightdue, for instance, to near-field interaction with the NWs, contributionsgiven by polarization-dependent light scattering by the NWs cannotbe completely ruled out. As discussed previously,9 polarization modulation measurements require us to modelthe behavior of the entire optical system and to account for the residualoptical activity of its components, including the optical fiber probe,eventually leading to an offset in γ. Since this offset remainsconstant during the SNOM scan, it can be determined and subsequentlysubtracted by performing reference measurements on bare substrates.The correct operation of the entire system is assessed through measurementsof polarizing materials oriented along different directions.Thenanoscale optical anisotropy of the nanocomposite fibers is investigatedby polarization-modulation SNOM. Details of this technique are reportedin our previous study.33.144 During extension and retraction, the mobility of a chain is constrainedby the potential induced by nearby chains and is thus confined withina tubelike volume, a confining tube . The polymer network is stretched affinelywith the flow, resulting in relative extension of chains defined byε = L/Lmax, where L is the primitive path of a chain . Thus, the chain extends ata rate of ε∇v due to affine convection.The ensuing net tension force in a stretched chain is f(ε) – f(ε0), where f(ε) is the normalized (dimensionless) elastic tensionat an extension ε, and f(ε0) is the tension at equilibrium (at rest). This force tends to retractthe chain through the tube back to equilibrium, within the Rouse relaxationtime τ. Thus, the chain retracts at a rate of −[f(ε) – f(ε0)]/τ due to relaxation. Combining the stretching and retractionrates, the rate of the relative extension is given by (details inSupporting Information Section S1):10The concepts of confiningtubes, chain reptation, and chain stretchingand retraction have been widely used in modeling the rheology of semidiluteentangled polymer solutions under extensional flow.52 typically within the range of s∼103–105 s–1. When the electrospinning jet reaches the steady state, the constantvelocity gradient is ∇v = s. Furthermore, since stretching and retraction reach equilibrium,the time derivative of the relative extension is zero. Consequently, N), the strain rate must be very high to reach substantial stress,and vice versa. In other words, the stress is the result of competitionbetween two time scales, that of the jet strain (s–1) and that of the chain relaxation (τ). The term sτ represents the instantaneous stiffness of the polymer liquid.The velocity gradient of the jet stream is fairly constant in aprocess,53 We use the followingapproximation for the elastic retraction force1048 see Supporting Information Section S1). Substituting the elastic force expression into 0 yield ε ≈ ε0(1 + sτ/3) for low sτand ε ≈ 1 – 3/sτ for high sτ (see Supporting Information Section S4). The calculation of the relaxation time τand initial extension ε0 for given polymer and solutionproperties is provided in Supporting Information Section S2.The network extension induced by the flow generates elastic tensionin the chains. The elasticity of a polymer chain is entropic, andtherefore, its tension is linear at small extension but rises sharplyat high extension associated with less probable chain conformations.Thus, the chain extension dependence of the tension force is nonlinear,and several approximations are known.33 The crowdingof chains near a surface gradually decays at a characteristic distancefrom the surface, in the order of the polymer network correlationlength. In other words, chains near the boundary are confined by thesurface in one of the lateral directions due to adsorption and thereforeessentially tend toward a two-dimensional conformation. The correspondingphysical properties in the 2D condition are marked by an overbar accent.The expression for the elastic tension force changes to f̅ ≅ 2ε̅(1 – ε̅0)/(1– ε̅), where the prefactor 3  for low sτ and ε̅≈ 1 – 2/sτ̅ for high sτ (see Supporting Information Section S4). These extended chain conformations are partiallyfrozen upon nanofiber solidification because of the rapid solventevaporation characteristic of electrospinning.Scaling approximationsat small ε̅When a polymerchain is dissolved in a solution and is free ofexternal loads, it assumes a compact coil shape. The extension expressionsin 10 By using s/sc = sτ/3 and s/s̅c = sτ̅/2, respectively.The critical strain rate determines such a transition:at low strainrates, the equilibrium between chain stretching and retraction issatisfied by small extensions; however, at high strain rates, theretraction stress must be high to balance stretching, leading to highextensions. High chain extension induces a high molecular alignment,which in turn enhances the material properties in the stretching direction.33 resulting in a localincrease in relaxation time (see the concentration dependence of τin Supporting Information Section S2).The effect would be a left shift of the adsorption limit in 0 ≅ε0. Both approximations have a minor effect on themolecular orientation because (i) at small extension, the orientationis dominated by the adsorption, regardless of the initial extensionor relaxation time, and (ii) at large extension, the relative effectof the small initial extension or relaxation time is negligible. Accordingly, Throughout our calculations, we use the approximation τ̅≅ τ, based on the assertion that the friction experiencedby a retracting chain is similar in both 2D and 3D conditions. Tobe exact, near a filler boundary, the monomer concentration slightlyincreases due to adsorption,3.2s, the chain relaxationtime τ and extension at rest ε0 are the variablesthat determine the degree of chain extension. A higher strain rateresults in a higher tension and extension. Similarly, a long relaxationtime slows down stress relieving, resulting in a higher extension.In the case of a polymer blend  between defects, eachbead having a diameter of 1.2 nm , is about 13 times shorter in MEH-PPVthan in PVP; hence, the MEH-PPV critical strain rate, scM, is about 13 times higher than that of PVP, scP, as seen in The two-polymer species differ significantly in their conformation.While the backbone of a PVP chain is composed of a sequence of singlecovalent bonds and is therefore flexible, the MEH-PPV chain is composedof alternating single and double covalent bonds (conjugated bonding),making it highly rigid in bending. However, inherent bonding defectsin the MEH-PPV partially substitute rigid conjugated links with flexibletetrahedral links, making the polymer chain semiflexible with longrigid segments (Kuhn monomers).f 1.2 nm 1. The exVP chain 1d. Simils < scP, where both polymers are only negligibly stretched; (b) the regionbetween the critical strain rates of PVP and MEH-PPV, scP < s < scM, where PVP is stretched significantly, whereas MEH-PPV isnot; and (c) the region above the critical strain rate of MEH-PPV, s > scM, where both polymersare stretched significantly. In the latter region, the high strainrate leads to rapid separation between entanglement nodes, resultingin the disentangling of chains from the network. While the PVP longchains may still be entangled in a network, maintaining the integrityand contiguousness of the network, the MEH-PPV chains may be fullydisentangled from the network because of their shorter length andlesser extension.57 For example, as theprimitive length of the PVP in that region is about an order of magnitudelonger than that of MEH-PPV, at the instant MEH-PPV disentangles (2nodes at its edges), PVP is still entangled in 10 nodes. The MEH-PPVchains keep moving with the network and are extended by the jet strainrate as expressed in the model in In terms of the relative extension,the PVP/MEH-PPV polymer blendhas three different regions: (a) the region below the critical strainrate of PVP, 3.3N successive Kuhn monomers of length b,placed on a 3D Cartesian lattice.32 Givena force iF acting in direction i, the energy associated with a monomer aligned in thatdirection is ibF. The corresponding statisticalBoltzmann factor is eibF/kBT–, where kB is the Boltzmann constant, and T isthe absolute temperature. The exponent if = ibF/kBT is the normalized tension force in direction i. Thus, the probability that a monomer will align in direction i is given by10y axis), the extensionalforce is yf = f(ε),whereas in the lateral directions, the force is that of a networkat rest, xf = zf = f(ε0). We ignore the effectof radial compression associated with longitudinal extension becauseit can be considered as negligible (see Supporting Information Section S3).Thus far, we have expressed the relative extensionof polymer chains,both those far from a boundary and those near it, in terms of therelative strain rate of the electrospinning jet. We now turn to estimatingthe molecular orientation associated with the extended conformationof chains. A linear polymer chain can be expressed as a sequence of y = Py+ – Py–, andthe initial extension is obtained by substituting yf = f(ε0). Using Hermans’orientation parameter, N segments of a chain, i± is the angle ofa segment with respect to the longitudinal axis. As in the Cartesianlattice y± = 0, we obtainThe relative extension isgiven by ε = εSection S3), one has0 isgiven in Supporting Information Section S2 and ε by 0 yield O ≈ ε02sτ for low sτ and O ≈ 1 – 9/2sτ for high sτ (see Supporting Information Section S4).Using z, and the probability of alignment in that direction isnull, that is Pz± = 0, and the terms containing zf are removed from x and y directions. The orientation in terms of theextension for this case is (see details in Supporting Information Section S3)0 ≅ ε0 isgiven in Supporting Information Section S2 and ε̅ by if. Scalingapproximations at small ε0 yield O̅ ≈ (1 + 3ε̅02sτ̅)/4 for low sτ and O̅ ≈ 1 –3/sτ̅ for high sτ(see Supporting Information Section S4).Near a boundary, such as thesurface of an NW, monomer alignmentperpendicular to the surface is precluded due to adsorption of monomers.Since the NWs are assumed to be aligned with the flow , V is the solution velocity, and l is theNW half-length; (ii) the acceleration force scales as ρd2lV̇, where ρ =7.2 g cm–3 is the NW density, d is its diameter, and V̇ = V∇V is the solution acceleration; the ratiobetween the forces is η/ρd2∇V∼106, meaning that thehydrodynamics is predominant.An NW in the electrospinningflow moves at the same velocity of thesolution near the NW center. This can be verified by comparing thehydrodynamic and acceleration forces acting on the NW: (i) the hydrodynamicforce scales as ηy from the NW center, v = sy , 2s atthe top (y = – l), and 0at the bottom (y = l).This prediction is an upper limit for the change in the strainrate, as the velocity difference between the chains adjacent to theNW and those far from it occurs across the fluid boundary layer. Ifthe boundary layer is larger than the scale of a typical chain, thechain will sense only a fraction of the velocity difference and hencea fraction of the strain rate change. The boundary layer thicknessis inversely related to the fluid velocity, and therefore, a thickerlayer tends to form near the NW center, resulting in a lower strainrate, and a thinner layer forms near the ends, resulting in a higherstrain rate. As a complete fluid analysis is beyond the scope of thecurrent study, we use the following simplified upper-bound expressionfor the strain rate along an NW boundary3.5x, y space, where x isthe radial distance from the NW boundary and y isthe axial downstream distance from the NW midpoint  applies onlywhen |xe couldpossibly be in the order of the polymer network correlation length b/φ,32 or the NW half-length l, both of order 102 nm. However, xe is not necessarily the same for adsorption and drag,and in such a case, xe with each contribution. For simplicity’ssake, the following maps assume the same extinction distance. Thelongitudinal extinction distance ye couldbe in the order of the NW length or a fraction of it.The radial extinction distance O is depicted in s/sc, for the initial extensionof MEH-PPV from The orientationmapping function As noted, beyond the NW ends, these effects are assumed to decayexponentially. Thus, at low strain rates, the orientation dominatedby adsorption decreases gradually beyond both NW ends to the low valueof the unperturbed flow. By contrast, at high stain rates, above theNW top end, the orientation enhanced by dragging decreases graduallytoward that of the unperturbed flow, whereas below the bottom end,the orientation reduced by restraining increases gradually towardthat of the unperturbed flow. One should keep in mind that these mapsrepresent upper-limit orientation values and should therefore be regardedmerely as trends to be supported by experiments , raising bothε0 and τ (equations in Supporting Information Section S2) and consequently the extension andlongitudinal molecular orientation. At the same time, the polymernetwork near the nanofiber boundary is extended in the radial direction,causing a radial molecular orientation. These effects are substantiatedtheoretically62 as well asexperimentally52 byX-ray imaging of jets, near-field optical imaging of nanofibers, andAFM elastic modulus measurements at nanofiber cross sections. Theycan also be observed by the optical measurements of nanofibers inthe current study . The jet longitudinal strain rate 3.6v0 (feed rate), the electric field intensity E, andthe solution concentration φ. The parametric dependence of therelative strain rate scales as 10 The other parameters include the monomer length,injector internaldiameter, solution electric conductivity, solvent viscosity, and degreeof polymerization. The exponents of φ for a good solvent (−2.96)and for a θ-solvent (−2.89) are similar, implying lowdependence on solvent quality.The relative strain rate and,consequently, the degree of chain extensionand molecular orientation depend on the electrospinning conditions.A complete analysis of all parameters involved in the electrospinningprocess is highly complex. We therefore focus on the readily tunableparameters—the jet exit velocity Section S4), the orientation far from an NW scales asO signifies the distance of the orientation from the possiblemaximum (O = 1). At low strain rates, the orientationscales as O ∝ s/sc. We see that the orientation increases when the electric field isincreased and when the feed rate or concentration is decreased. Thedifferent power exponents indicate that coarse tuning can be conductedby modifying the concentration, and moderate tuning can be conductedby modifying the electric field, whereas fine tuning is achieved bymodifying the feed rate. It should be noted that, in order to ensurejet formation, continuity, and contiguousness and to prevent jet breaking,the electrospinning process parameters should be kept within the physicalbounds of the process by remaining within the combined working rangesof the feedrate, electric field, and solution concentration.57Using a scaling approximationfor high strain rates ,close to zero, with a standard deviation of 5 degrees.The fluorescence micrographs 7a,b show4.2Figure S1).3 NWs  are lower in the NW vicinity than further away from it  by a factor of 1.7. In 58Figure S2 in the Supporting Information.This is indicative of an average increase of the alignment of theMEH-PPV macromolecules caused by the NWs.The presence of NWs impacts the nanoscalelocal arrangement ofthe MEH-PPV macromolecules and determines, on average, a local spatialvariation of the dichroic ratio. In fact, the presence of the NW leadsto an average increase of the absolute value of γ (by a factorof about 2) at the fiber central axis, with MEH-PPV macromoleculesaligned along the fiber length, as can be observed by comparing theline profiles of γ along directions parallel and perpendicularto the NW length .The results obtained from the theoreticalmodel we developed arein line with the experimental findings. Let us start with the exampleof y/l = – 1), both the experimental and theoretical molecular orientationsrise sharply above the values far from the NW by a factor of about2, owing to the combined effect of the higher drag at the fiber topand the adsorption of polymer chains near an object. Toward the NWmidpoint (y/l = 0), both the experimentaland theoretical molecular orientations decrease moderately as a resultof the diminishing strain rate and drag but are still higher thanvalues obtained far from the NW. At the midpoint, the drag effectvanishes, whereas the adsorption effect still enhances the orientation.Near the top end of the NW , the orientationin this example is about 0.2, implying a higher relative strain rate,likely expressed by the theoretical curve of s/sc = 1.5 in y/l = 1), where thelow strain rate and drag counteract the adsorption. This is indeedobserved in the black curve in s/sc ≤ 1), there is no negativepeak in that region because the effect of adsorption is far greaterthan that of drag.These trends can be observed in the example of the Supporting Information, S2b. The peak experimental value is about 0.3,whereas the corresponding theoretical values are about twice as high.The major reason for this difference is likely the partial relaxationof the polymer network after the jet reaches the collector, when furtherstretching ceases, while the fiber is not yet fully solidified.As already noted, the theoretical analysisrepresents upper-limitorientation values. We find evidence of this in the value of Herman’sorientation parameter derived from the dichroic ratio in 53 NWs. Our aim is to rationalizeand ultimately enhance the orientation of conjugated polymer chains,a condition critically important for improving the polarization degreeof the light emitted by the hybrid nanofibers. It is shown that thepolymers undergo a stretch transition at a critical jet strain rate,leading to the corresponding high molecular orientation. Adsorptionof chains near the boundary of an NW causes further orientation alongthe jet axial direction. Furthermore, the hydrodynamic drag inducedby an NW on the flow affects the local strain rate, with matchingvariation in local orientations. These effects are mapped and tunedto the electrospinning conditions.The molecular orientation ofan electrospun MEH-PPV conjugatedpolymer, blended with a PVP polymer, is studied theoretically, focusingon the extension caused by electrospinning and the local adsorptionand drag induced by WO3 nanofibers, imaged bypolarized SNOM. The mapping of the optical dichroic ratio of the MEH-PPVdemonstrates the molecular anisotropy caused by the NWs. These measurementsalso unveil an increased molecular alignment of the polymer moleculesalong the fiber axis in the vicinity of the NWs, supporting the predictedeffect of adsorption. Furthermore, the dichroic ratio is not distributeduniformly along the NW length, indicating the predicted effect ofhydrodynamic drag.These predictions are supportedby experimental evidence derivedfrom electrospun MEH-PPV/PVP/WO3 NWs with the MEH-PPV conjugated polymercould be advantageousfor the optical properties of the nanofiber, potentially improvingits ability to transport light excitations as a result of molecularalignment. Further research is envisioned using higher resolutionoptical scanning of nanofibers to map the molecular orientation, characterizationof the optical properties of these fibers, and tuning of the electrospinningmaterials and process conditions.These synergetic results demonstrate thatthe mixing of WO


Historically, cancers presenting with their main tumor mass in the ovary have been classified as ovarian carcinomas  and treated with a one-size-fits-all approach. Over the last two decades, a growing molecular understanding established that ovarian carcinomas consist of several distinct histologic types, which practically represent different diseases. Further research is now delineating several molecular subtypes within each histotype. This histotype/molecular subtype subclassification provides a framework of grouping tumors based on molecular similarities for research, clinical trial inclusion and future patient management.POLE-mutated/mismatch repair-deficient/no specific molecular profile/p53-abnormal) with the same prognostic stratification as their endometrial counterparts. Although methylation analyses and gene expression and sequencing showed at least two clusters, the molecular subtypes of CCCs remain largely elusive to date. Mutational and immunohistochemical data on LGSC have suggested five molecular subtypes with prognostic differences. While our understanding of the molecular composition of ovarian carcinomas has significantly advanced and continues to evolve, the need for treatment options suitable for these alterations is becoming more obvious. Further preclinical studies using histotype-defined and molecular subtype-characterized model systems are needed to expand the therapeutic spectrum for women diagnosed with ovarian carcinomas.The phenotypically informed histotype classification remains the mainstay of ovarian carcinoma subclassification. Histotypes of ovarian epithelial neoplasms have evolved with each edition of the WHO Classification of Female Genital Tumours. The current fifth edition (2020) lists five principal histotypes: high-grade serous carcinoma (HGSC), low-grade serous carcinoma (LGSC), mucinous carcinoma (MC), endometrioid carcinoma (EC) and clear cell carcinoma (CCC). Since histotypes arise from different cells of origin, cell lineage-specific diagnostic immunohistochemical markers and histotype-specific oncogenic alterations can confirm the morphological diagnosis. A four-marker immunohistochemical panel (WT1/p53/napsin A/PR) can distinguish the five principal histotypes with high accuracy, and additional immunohistochemical markers can be used depending on the diagnostic considerations. Histotypes are further stratified into molecular subtypes and assessed with predictive biomarker tests. HGSCs have recently been subclassified based on mechanisms of chromosomal instability, mRNA expression profiles or individual candidate biomarkers. ECs are composed of the same molecular subtypes ( The subclassification of ovarian carcinomas is now based on a hierarchical approach; the first step is to subclassify based on traditional histopathological phenotypes into histotypes. Histotypes are considered different diseases based on the cell of origin, molecular alterations, clinical behavior and management ,2. Precitubo-ovarian high-grade serous carcinoma . ,23. 3,23The specific use of ancillary IHC markers depends on the diagnostic considerations, which can be confirmatory  might be performed in rare high-grade cases that cannot be reliably classified. Serous carcinomas with moderate (grade 2) nuclear atypia may be subject to p53 IHC to distinguish HGSC from LGSC. This has only become possible after IHC optimization to accurately predict TP53 mutation status [The differential diagnostic approach between two entities is divided into first-line panels, which solve most of the cases and are sufficient if the morphological context is compatible, and more extensive second-line panels, which may be reserved for cases with phenotypes contradicting the first-line panel, unexpected first-line panel results or other unusual constellations. WT1 is the most important marker that is diffusely expressed in almost all HGSCs and LGSCs and virtually absent in almost all CCCs and MCs. However, it can be expressed in 10–15% of ECs ,23. Thern status ,53. The n status . The besn status . MCs aren status . There iThere are many ways to subclassify HGSCs . HGSCs aCCNE1 high-level amplification (>eight copies) [BRCA1/2 mutations in patients, which could be at least partly due to a better response to platinum–taxane therapy [Numerous studies have developed prognostic mRNA signatures for HGSC , but a r copies) ,63,64,65 copies) . Another therapy . Moreove therapy .BRCA1-associated tandem duplications, BRCA2-associated interstitial deletions, CCNE1-amplified associated fold-back inversions and CDK12-associated tandem duplications, and they showed that these are associated with different mechanisms of immune resistance, explaining the disappointing results in recent immune checkpoint inhibitor trials that recruited thousands of women diagnosed with HGSC [HGSC is the prototype of a chromosomally unstable cancer. Brenton and colleagues defined seven distinct copy number signatures, each associated with a different mechanism of chromosomal instability . Shah anith HGSC ,71.BRCA1/2 mutation status to commercial HRD tests [CCNE1 given its mutual exclusivity to BRCA1/2 germline mutations as shown by Bowtell and colleagues [The breakthrough for the treatment of HGSC was the recent approval of PARP inhibitors as a standard of care . HoweverRD tests . Even thRD tests . It remaRD tests ,75, partRD tests . It may lleagues ,65,76.POLE mutated, MMRd, no specific molecular profile (NSMP) and p53 abnormal (p53abn)) with the same prognostic stratification as their endometrial counterparts [POLE mutation (POLEmut) have the most favorable prognosis, while patients with p53abn tumors can expect an aggressive disease course. MMRd and NSMP are associated with an intermediate prognosis. This stratification remained significant in uni- and multi-variable analyses when restricted to low-stage (defined as stages I–IIA) cases and provided better stratification than a histologic grade, providing further evidence that grading may eventually be replaced by molecular determinants. In contrast to the endometrium, however, the group of NSMP is substantially larger (73% versus 56%), requiring further stratification. The most promising biomarkers, which have only been assessed outside the context of molecular subtype thus far, are PR and CTNNB1, with the latter being the most commonly mutated gene in ovarian EC [One large study and other smaller studies have established that ovarian endometrioid carcinomas are composed of the same four molecular subtypes  and mesonephric-like adenocarcinomas (NSMP) in the group of endometrioid carcinomas. Excluding those and p53abn, a diagnosis of low-stage endometrioid carcinoma of other molecular subtypes without loss/reduced PR expression represents the best group for surveillance.ARID1A mutations as the most common molecular alteration in CCC [TP53 mutations and cluster 2 by co-occurring ARID1A/PIK3CA mutations and Asian ancestry) [ERBB2 amplifications occur in 7%–14% of CCCs, making it a good candidate for inclusion into basket trials [Advanced clear cell carcinoma remains a therapeutic dilemma. Huntsman and colleagues discovered n in CCC , but then in CCC . ARID1A,n in CCC . Recent ncestry) . Gene exncestry) . Individncestry) ,91. Nevencestry) , and thet trials ,93.Clinical trials and case reports suggest immune therapy; however, biomarker development to predict response to checkpoint inhibitor therapy has been challenging without consistent predictors  . MMRd wiKRAS, NRAS, BRAF) ~ USP9X mutations ~ NSMP [Data from a large study integrating mutational data from targeted sequencing and IHC in LGSC suggest five molecular subtypes, and, listed in order of decreasing aggressiveness, they are CDKN2A IHC alteration > PR loss/high fraction of genome altered > MAPK pathway mutations  of y events . The proerations ,103. In ot occur . The mosmutation . DespiteThe phenotypically informed histotype classification remains the mainstay of ovarian carcinoma subclassification. The histotype classification is particularly robust because histotypes arise from different cells of origin, allowing for cell lineage-specific diagnostic ancillary IHC markers in combination with histotype-specific oncogenic alterations. Ancillary IHC dramatically improves the precision of diagnostic histotyping.CCNE1 and CDK12) is a pragmatic strategy for biomarker test development within clinical trials. While our understanding of the molecular composition of ovarian carcinomas has significantly advanced, the need for treatment options suitable for these alterations becomes more and more obvious. To expand the therapeutic spectrum, preclinical studies require histotype-defined and molecular subtype-characterized model systems [The phenotype–genotype correlation has its limitations when it comes to molecular subtyping within histotypes. Phenotypes can direct certain tests , but mos systems .


Bovith recene tuberculosis (bTB) continues to be a pathogen of concern in several countries globally. Analysis of areas that have higher incidences of bTB outbreaks has demonstrated how risk is not equally distributed, and local data collection, analysis and participatory engagement is required to develop tailored approaches. The Burren, an internationally important heritage landscape, has been an area of higher bTB incidence for many years in Ireland, and owing to its unique geology and farming heritage a survey was developed to engage with local farmers to gain greater insight into farming practices and bTB control to inform tailored approaches. The survey gathered data on the farm and animal management approaches being used within the Burren, including local farming techniques like the use of “winterage” (grazing exposed limestone dominated uplands). Thematic analysis of free text responses was undertaken. Quantitative data were then explored using statistical models to assess associations with recent (<3 years) self-reported bTB breakdown risk. There was a high number of responses demonstrating a high degree of willingness to engage on the issue. Thematic analysis suggested that wildlife and its management (culling and vaccination), testing quality, and its impact on the bTB scheme, and pessimism around eradication were important themes. Statistical analysis suggested that increasing bTB risk was primarily related to increasing herd-size and the percentage of herd owner's land inaccessible to those attempting to locate badger setts. There was less evidence for associations relating to the amount of time, or which season , farmers utilized “winterage”. The results of the study will feed back directly to local bTB management plans and further stakeholder engagement and is an exemplar for local tailoring of national control measures in situations of high incidences of bTB outbreaks in particular areas. Mycobacterium bovis, with advanced infection resulting in chronic respiratory disease characterized by granulomas in affected tissues. This disease is a persistent problem having significant animal health and welfare consequences for cattle industries (Bovine tuberculosis (bTB) is caused by dustries  and posidustries  and thisdustries . Stakehodustries . The levdustries .Meles meles is a reservoir host of infection known to spillback infection to cattle  or number of cattle (question 8), the options were grouped numerically in order of size. In the ranking style question, (question 11), the participants had to rank each of the options; with 1 being what they felt the most important factor was. The method used for scoring these responses was to assign a reverse score, i.e., in a question with four options the most important factor, ranked by a farmer at number one, was given a score of four. The weighted average was then used to determine the rankings. It was made compulsory to provide a response to each question before one could progress to answering the next question. The final question was an open question, asking participants to “provide any further comments they had regarding bTB in the Burren”. Qualitative research used in open questions asks participants “to describe their experiences in ways that are meaningful to them”  team within the Irish Department of Agriculture, Food and the Marine (DAFM), are approved through a Departmental governance and oversight process. Informed consent was achieved by providing an information section at the beginning of the survey. This section explained the aims of the study, assured participants that their participation was voluntary and that their confidentiality would be protected. Respondents were free to choose whether to consent to participate in the study by selecting the option in the first question. Quotes from the qualitative data are presented anonymously in this manuscript.SurveyMonkey was used to generate rating scales for the ranking style question, as in previous research including Sayers et al. . Data frThe twentieth and final question of the survey asked participants to “provide any further comments you have regarding bovine TB in the Burren”. One hundred and sixty participants who had completed all the other quantitative questions ended the survey without providing any further comments. A total of 110 responses were received for this question (representing 37% of the respondents who had consented to participate in the study). The first step in the analysis of these free text responses was to identify any unusable replies, for example, where the respondent had indicated that they had no further comment. Six responses were removed at this stage, leaving 104 substantive responses for further analysis. This question produced a significant volume of qualitative data, coming to a total word count of 4,764 words.Each of the individual response was read and assigned a tag, or multiple tags where appropriate, categorizing the responses as falling under specific themes. It is not claimed that the list of themes used to categorize the qualitative responses is an exhaustive one, rather the themes cover those topics which could be clearly defined and which arose in multiple responses. Twenty six of the responses to this question referred, in whole or partially, to issues that could not be categorized clearly within these chosen themes. Certain select quotes were be used to illustrate some of these miscellaneous topics in this paper.In order to explore correlates of bTB risk, all herds were classified as either being recent bTB positive or negative, based on self-reporting from participants. All herds were classified as “positive” if herd owners reported that their herds experienced as bTB breakdown within the last 3 years. All other herds were classified as “negative” for recent bTB breakdown.All categorical variables allowed for only one category per herd. Where a farmer selected two options for herd type, only one was used in the model. The response was always taken as the option with the subjective higher bTB risk, given previous research from Ireland e.g., , 15–17].–17.15–17Throughout, the binary outcome for models was used to assess whether there were any associations with putative risk factors, based on survey responses. Logit regression models were used to model these associations. All independent variables were categorical factor variables. Each putative risk factor was assessed for [unconditional] association with the outcome using univariable models. Variables were then offered to multivariable models, with model building completed in two ways. Firstly, parsimonious models were selected using automated backwards, forwards, and stepwise selection procedures (using Stata's STEPWISE function). In addition, as there were many independent variables and a relatively small dataset, a multi-model comparison was also undertaken using Akaike's Information Criteria, corrected for small sample size (AICc). The premise being that the “true” model may be one of a set of candidate models , 19. Eacn = 303). The majority of farms in the Burren were classified as suckler farms (64%) . Other farm enterprises included suckler herds where replacements are purchased (19%) and herds where cattle were purchased and fattened for sale (14%). Less than 10% of farms were dairy enterprises.Initially, 280 participants completed the first survey questions, meaning twenty participants dropped out immediately after consenting to take part  of farmers either had their last bTB outbreak over 10 years ago or have never had a breakdown. Approximately 30% (83/280) of farmers had bTB detected in their herd within the past year, while a further 13% of Burren farmers last had bTB in their herd between 1 and 3 years ago (35/280). This meant for our modeling outcome, 118/280 (42%) herds were classified as a positive case .The unconditional associations with this outcome, when p<0.1, are presented in The composition of the ten highest ranked models are presented in Based on the multi-model comparisons, the inclusion probability for each variable is presented in The top ranked model is presented in p = 0.034) and 19.71 times the odds  of recently having a breaking down, respectively, relative to herds with 10% or less of their land inaccessible, while controlling for herd size.The top ranked model suggested that herds where farmers reported that 31–50% or over 50% of their land was inaccessible for those seeking to locate badger setts had 3.93  and 3.08  times the odds of recently experiencing a breakdown, respectively, relative to herds with ≤30 animals. The predicted increase in risk by herd-size and badger sett inaccessibility percentage is presented in Large herds of 51–100 animals and very large herds of >100 animals were 2.17 , most of which mentioned the difficulty of maintaining boundary walls due to feral goats, water sources (n = 3), and cattle movement (n = 3) were the next most frequently mentioned topics.Biosecurity  and several significant associations were identified. The results will inform improved policy development by DAFM, particularly in relation to bTB risk communication and stakeholder discussions on how best to engage with herdowners at the bTB Forum.Our statistical exploration of the survey data suggested that there was an association between recent bTB breakdown and the response to the question “what percentage of your entire farm is inaccessible (scrub or hazel) to locating badger setts?” There was a trend toward increasing risk of recent bTB breakdown and the percentage of farm inaccessible to setts due to habitat. At face value, this result could suggest that herds with more suitable habitat for badgers may be at greater risk for bTB breakdown; indeed, previous research has suggested that there is an association between metrics of badger density and breakdown risk . InaccesIn addition to the management of wildlife, several respondents made comments on the overall management of the bTB eradication programme. Some voiced concerns regarding demands created by frequent testing of animals for bTB and this reflected fatigue with the eradication programme. This finding is similar to that of Robinson  where itThe size of cattle herds has consistently been a risk factor for bTB breakdowns in Ireland and elsewhere when using large national-scale datasets e.g., , 28, 29], 2928, 2A recent study found that in addition to herd size, the number of parcels of land used can be associated with increased risk, but not metrics of intensity based on nitrogen loading, with breakdown duration . Herd siGrazing winterage habitats is a unique farming approach confined to the Burren region, and is a result of the unique geology of the region . The expThis is a relatively small sample size but there was a good response. Furthermore, it was representative of the population in the Burren, which was the subject of this study. The survey was anonymous which prevented us from verifying the herd's bTB infection history as herd identifier and location was not obtained in the survey. However, this was necessary for data protection purposes and may have encouraged the farmers to express their views more freely.We sought to analyse risk factors through participatory engagement with farmers and get their perspectives on bTB eradication in a specific region in Ireland. The level of engagement in the survey suggests that farmer stakeholders are very keen to be involved in and help with the control and eventual eradication of bTB in their area. The responses to the qualitative questions indicate concerns around wildlife management, fatigue with disease eradication programme, the quality of testing applied in the bTB programme and general pessimism around eradication of the disease. There was some of evidence of a mismatch between perception of risk by farmers, and actual risk relating to feral goats, given available data suggesting there is no M bovis circulating within that population. Statistical analysis suggested increasing bTB risk was primarily associated with increasing herd-size and the percentage of herd owner's land inaccessible to locating badger setts. The exploration of the utilization of “winterage”, a unique farming approach to the Burren, showed an indication of a potential association between recent bTB risk and amount of time winterage was used during the winter season and also whether winterage was used in summer months. Further research would be required to examine this latter finding in more detail. The results of the study can be utilized in local bTB management plans so that measures are tailored to the needs of this specific location.The original contributions presented in the study are included in the article/AC drafted the manuscript. JM designed the study and administered the questionnaire. AB carried out the statistical analysis. FF and CM provided local input into the design and distribution of the survey. ER came up with the concept. DB oversaw the overall project. All authors have input into the production of the manuscript.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.