Discourses on Positive Animal Welfare by Sheep Farmers and Industry Actors: Implications for Science and Communication.

This research examines how sheep farmers and industry actors in the United Kingdom (UK) understand and conceptualize what animal welfare scientists term 'positive animal welfare'. It explores their awareness of the concept, and how they interpret it using a qualitative approach. Participants were recruited using a snowballing, purposive sample approach, resulting in 25 sheep farmers and 11 industry actors (veterinarians, farming organizations, advisors, and supply chain) being interviewed. To collect data, a combined approach involving semi-structured interviews and a facilitated workshop were used between April 2021 and March 2022. Data were then thematically analyzed using a hybrid of inductive and deductive coding process. The findings suggested that the perceptions of farmers and industry actors in the study regarding positive welfare differ from contemporary academic discourses. Overall, around 7 of the farmers equated positive welfare with "positive stockmanship", while six of them expressed "good animal welfare" definitions associated with the Five Freedoms. In contrast, most industry actors (6) expressed interpretations associated with high welfare standards (going above minimum recommended practices) and positive mental experiences (3). Emerging discourses revealed the link between self-identity, social identity and what positive welfare is, the importance of knowledge exchange, and the need for practical indicators through language rephrasing. There is a clear need to enhance and improve knowledge dissemination strategies, particularly in the UK, where much research is being conducted on positive animal welfare.

Specific T-cell subsets have a role in anti-viral immunity and pathogenesis but not viral dynamics or onwards vector transmission of an important livestock arbovirus.

Introduction: Bluetongue virus (BTV) is an arthropod-borne Orbivirus that is almost solely transmitted by Culicoides biting midges and causes a globally important haemorrhagic disease, bluetongue (BT), in susceptible ruminants. Infection with BTV is characterised by immunosuppression and substantial lymphopenia at peak viraemia in the host. Methods: In this study, the role of cell-mediated immunity and specific T-cell subsets in BTV pathogenesis, clinical outcome, viral dynamics, immune protection, and onwards transmission to a susceptible Culicoides vector is defined in unprecedented detail for the first time, using an in vivo arboviral infection model system that closely mirrors natural infection and transmission of BTV. Individual circulating CD4+, CD8+, or WC1+ γδ T-cell subsets in sheep were depleted through the administration of specific monoclonal antibodies. Results: The absence of cytotoxic CD8+ T cells was consistently associated with less severe clinical signs of BT, whilst the absence of CD4+ and WC1+ γδ T cells both resulted in an increased clinical severity. The absence of CD4+ T cells also impaired both a timely protective neutralising antibody response and the production of IgG antibodies targeting BTV non-structural protein, NS2, highlighting that the CD4+ T-cell subset is important for a timely protective immune response. T cells did not influence viral replication characteristics, including onset/dynamics of viraemia, shedding, or onwards transmission of BTV to Culicoides. We also highlight differences in T-cell dependency for the generation of immunoglobulin subclasses targeting BTV NS2 and the structural protein, VP7. Discussion: This study identifies a diverse repertoire of T-cell functions during BTV infection in sheep, particularly in inducing specific anti-viral immune responses and disease manifestation, and will support more effective vaccination strategies.

Stakeholder Challenges and Opportunities of GPS Shock Collars to Achieve Optimum Welfare in a Conservation or Farm Setting.

Virtual fences for livestock facilitated by a GPS shock collar (GPS-SC) and phone app were introduced to the UK in cattle herd trials in 2020. Technology which uses aversive shocks to control livestock movement on farms and in other settings poses a significant risk to livestock welfare. There are currently no welfare protocols in place in the UK to ensure the ethical use of GPS-SCs. The objective of this study was to understand how GPS-SCs were being used in practice in the UK and gather data to assist researchers and policymakers in the future research and development of a welfare protocol for the UK. We studied how the technology performs in terms of welfare challenges and opportunities, covering extensive livestock production, conservation settings, "rewilding", and regenerative farming practices, where the technology is currently being applied. Semistructured interviews were conducted with key stakeholders. In-depth interviews (n = 8) supported the previous literature that the use of GPS-SCs in restricted grazing settings poses a risk to animal welfare. This is due to the wavering virtual fence boundary line (which is affected by satellite movements), a lack of visual markers, and, in some "rewilding" and conservation settings, livestock keepers, which require training and support to enable optimal welfare in practice and prevent misuse of the technology. Results also indicated that there are opportunities for enhancing livestock welfare with GPS-SCs in very extensive farm settings, where targeted care can be facilitated by using the data to monitor and track livestock using GPS-SCs, and which can also prevent cattle injury or fatality through virtual pastures designed to protect livestock from hazards such as roads or bogs. Future research is needed to focus on minimising shocks in the training period and to better understand the value of visual electric fences in the training process.

Targeting foam cell formation to improve recovery from ischemic stroke.

Inflammation is a crucial part of the healing process after an ischemic stroke and is required to restore tissue homeostasis. However, the inflammatory response to stroke also worsens neurodegeneration and creates a tissue environment that is unfavorable to regeneration for several months, thereby postponing recovery. In animal models, inflammation can also contribute to the development of delayed cognitive deficits. Myeloid cells that take on a foamy appearance are one of the most prominent immune cell types within chronic stroke infarcts. Emerging evidence indicates that they form as a result of mechanisms of myelin lipid clearance becoming overwhelmed, and that they are a key driver of the chronic inflammatory response to stroke. Therefore, targeting lipid accumulation in foam cells may be a promising strategy for improving recovery. The aim of this review is to provide an overview of current knowledge regarding inflammation and foam cell formation in the brain in the weeks and months following ischemic stroke and identify targets that may be amenable to therapeutic intervention.

Positive Aspects of Welfare in Sheep: Current Debates and Future Opportunities.

The concept of positive welfare is an expansion of the traditional understanding that animal welfare is defined by minimizing stress, pain, suffering, and disease. Positive welfare shifts the animal welfare narrative from a focus on reducing negative experiences to proactively providing animals with opportunities to have positive experiences and feelings. The concept, although around for several decades, is in its infancy in terms of developing ways of assessing positive welfare on farms, especially in extensive systems, and there are challenges in the adoption of positive welfare practices and the monitoring of continuous improvement at the farm level. Using an iterative approach, this critical review aims to explore the extent to which positive welfare interventions and indicators are positioned and have been developed within the animal welfare literature for sheep. This paper critiques existing positive welfare indicators, such as choices in food and the physical environment, conspecific social synchronization, maternal bonds, intergenerational knowledge transfer, positive human-animal relationships, etc., as currently assessed by the 'good life framework'. It also reviews the characteristics of scientific measures for (positive) affective states in the current sheep literature and their potential contribution to understanding positive welfare states in sheep. In conclusion, this paper provides recommendations for future research regarding sheep welfare.

A "Good Life" for Dairy Cattle: Developing and Piloting a Framework for Assessing Positive Welfare Opportunities Based on Scientific Evidence and Farmer Expertise.

On-farm welfare assessment tends to focus on minimising negative welfare, but providing positive welfare is important in order to give animals a good life. This study developed a positive welfare framework for dairy cows based on the existing scientific literature which has focused on developing positive welfare indicators, and trialled a participatory approach with farmers; refining the framework based on their recommendations, followed by a vet pilot phase on farm. The results revealed that farmers and scientists agree on what constitutes "a good life" for dairy cattle. Farmers value positive welfare because they value their cows' quality of life, and want to be proud of their work, improve their own wellbeing as well as receive business benefits. For each good life resource, the proportion of farmers going above and beyond legislation ranged from 27 to 84%. Furthermore, barriers to achieving positive welfare opportunities, including monetary and time costs, were not apparently insurmountable if implementation costs were remunerated (by the government). However, the intrinsic value in providing such opportunities also incentivises farmers. Overall, most farmers appeared to support positive welfare assessment, with the largest proportion (50%) supporting its use within existing farm assurance schemes, or to justify national and global marketing claims. Collaborating with farmers to co-create policy is crucial to showcase and quantify the UK's high welfare standards, and to maximise engagement, relevance and uptake of animal welfare policy, to ensure continuous improvement and leadership in the quality of lives for farm animals.

The Social Construction of Narratives and Arguments in Animal Welfare Discourse and Debate.

Stakeholders can hold conflicting values and viewpoints, on what animal welfare is and how a good life is achieved and can signal different problems, or problematize specific aspects of farm animal welfare, and propose different actions or interventions within food supply chains. The aim of the study is to explore the contribution of narrative and argumentative discourse to the social construction and framing of animal welfare and its implications. The methodological approach in this research is composed of two phases with phase 1 being the foundational structured literature search in both academic and grey literature. Phase 2 was the analysis of the secondary data from the literature review to develop a synthesized iterative paper and in doing so develop a typology of five narratives: the 'farming as a business' narrative, the 'religion-based' narrative, the 'research, legislative and political based narrative', the 'higher welfare' narrative, and the "animal rights/power-based" narrative. Our findings demonstrate the contestation within the stakeholder discourse of the articulation of why farm animals should have a good life. Performance-related perspectives are rooted in the value-laden language and narratives that shape the arguments regarding notions of good and bad welfare; the emergent positioning of positive welfare for farm animals as well as how to achieve a good life in practice. The novel contribution of this review is the application of an explanatory word-language-discourse-person-situation-environment framework in this specific context to inform future research on animal welfare discourse analysis.

Murine precursors to type 1 conventional dendritic cells induce tumor cytotoxicity and exhibit activated PD-1/PD-L1 pathway.

The immediate precursor to murine type 1 conventional DCs (cDC1s) has recently been established and named "pre-cDC1s". Mature CD8α+ cDC1s are recognized for suppressing graft-versus-host disease (GvHD) while promoting graft-versus-leukemia (GvL), however pre-cDC1s have not previously been investigated in the context of alloreactivity or anti-tumor responses. Characterization of pre-cDC1s, compared to CD8α+ cDC1s, found that a lower percentage of pre-cDC1s express PD-L1, yet express greater PD-L1 by MFI and a greater percent PIR-B, a GvHD-suppressing molecule. Functional assays were performed ex vivo following in vivo depletion of CD8α+ DCs to examine whether pre-cDC1s play a redundant role in alloreactivity. Proliferation assays revealed less allogeneic T-cell proliferation in the absence of CD8α+ cDC1s, with slightly greater CD8+ T-cell proliferation. Further, in the absence of CD8α+ cDC1s, stimulated CD8+ T-cells exhibited significantly less PD-1 expression compared to CD4+ T-cells, and alloreactive T-cell death was significantly lower, driven by reduced CD4+ T-cell death. Tumor-killing assays revealed that T-cells primed with CD8α-depleted DCs ex vivo induce greater killing of A20 B-cell leukemia cells, particularly when antigen (Ag) is limited. Bulk RNA sequencing revealed distinct transcriptional programs of these DCs, with pre-cDC1s exhibiting activated PD-1/PD-L1 signaling compared to CD8α+ cDC1s. These results indicate distinct T-cell-priming capabilities of murine pre-cDC1s compared to CD8α+ cDC1s ex vivo, with potentially clinically relevant implications in suppressing GvHD while promoting GvL responses, highlighting the need for greater investigation of murine pre-cDC1s.

Emergence dynamics of adult Culicoides biting midges at two farms in south-east England.

BACKGROUND: Culicoides biting midges (Diptera: Ceratopogonidae) are biological vectors of livestock arboviruses that cause diseases with significant economic, social and welfare impacts. Within temperate regions, livestock movement during arbovirus outbreaks can be facilitated by declaring a 'seasonal vector-free period' (SVFP) during winter when adult Culicoides are not active. In this study we carry out long-term monitoring of Culicoides adult emergence from larval development habitats at two farms in the UK to validate current definitions of the SVFP and to provide novel bionomic data for known vector species. METHODS: Standard emergence traps were used to collect emerging adult Culicoides from dung heaps at two cattle farms in the south-east of England from June to November 2016 and March 2017 to May 2018. Culicoides were morphologically identified to species or complex level and count data were analysed using a simple population dynamic model for pre-adult Culicoides that included meteorological components. RESULTS: More than 96,000 Culicoides were identified from 267 emergence trapping events across 2 years, revealing clear evidence of bivoltinism from peaks of male populations of Culicoides obsoletus emerging from dung heaps. This pattern was also reflected in the emergence of adult female Obsoletus complex populations, which dominated the collections (64.4% of total catch) and emerged throughout the adult active period. Adult male C. obsoletus were observed emerging earlier than females (protandry) and emergence of both sexes occurred throughout the year. Culicoides chiopterus and Culicoides scoticus were also identified in spring emergence collections, providing the first evidence for the overwintering of larvae in dung heaps for these species. CONCLUSIONS: This study demonstrates continual and highly variable rates of emergence of Culicoides throughout the year. A lack of evidence for mass emergence in spring along with the ability to observe male generations highlights the need for complementary surveillance techniques in addition to light-trap data when investigating seasonality and phenology. Evidence was found of other vector species, C. chiopterus and C. scoticus, utilising cattle dung heaps as an overwintering habitat, further highlighting the importance of these habitats on farms.

Diversity and abundance of tabanids in Northern Spain.

Tabanids (Diptera: Tabanidae) are large haematophagous flies that cause both direct (by biting nuisance) and indirect (primarily by mechanical transmission of diseases) damage to host species. Research studies on this family have received little attention in some parts of Europe. Our aims were to characterise the species richness, abundance, and peak of activity of tabanid fly species in a region of Northern Spain. Home-made canopy traps, sweep nets, and Malaise traps were employed for the collection of tabanids across four cattle farms, two equestrian centres, and two golf courses during a 3-month period in the summer of 2020. A total of 300 specimens of 27 tabanid species belonging to eight genera were identified. The most prevalent species were Haematopota pluvialis (23.3%), Tabanus eggeri (20.0%), and Tabanus bromius (8.0%). The former species was recorded biting humans and therefore should be considered of relevance to public health. Tabanids were more diverse and abundant in scrubland and grazing pastures [relative abundance (RA) = > 10%; species richness (S) = 8-12; Shannon-Index (H´) = 1.5 - 2.1] compared to crop landscapes (RA = < 1%; S = 0-1; H´ = 0) according to canopy traps. The tabanid population dynamics was determined to be short, with the greatest abundance and diversity concentrated in mid-late July. This study updates the checklist of this Diptera group in the Northern Spain from nine known extant species to 31 species, providing the first data on the summer peaks of activity of tabanids for this region.

Regulatory Dendritic Cells Induced by Bendamustine Are Associated With Enhanced Flt3 Expression and Alloreactive T-Cell Death.

The growth factor Flt3 ligand (Flt3L) is central to dendritic cell (DC) homeostasis and development, controlling survival and expansion by binding to Flt3 receptor tyrosine kinase on the surface of DCs. In the context of hematopoietic cell transplantation, Flt3L has been found to suppress graft-versus-host disease (GvHD), specifically via host DCs. We previously reported that the pre-transplant conditioning regimen consisting of bendamustine (BEN) and total body irradiation (TBI) results in significantly reduced GvHD compared to cyclophosphamide (CY)+TBI. Pre-transplant BEN+TBI conditioning was also associated with greater Flt3 expression among host DCs and an accumulation of pre-cDC1s. Here, we demonstrate that exposure to BEN increases Flt3 expression on both murine bone marrow-derived DCs (BMDCs) and human monocyte-derived DCs (moDCs). BEN favors development of murine plasmacytoid DCs, pre-cDC1s, and cDC2s. While humans do not have an identifiable equivalent to murine pre-cDC1s, exposure to BEN resulted in decreased plasmacytoid DCs and increased cDC2s. BEN exposure and heightened Flt3 signaling are associated with a distinct regulatory phenotype, with increased PD-L1 expression and decreased ICOS-L expression. BMDCs exposed to BEN exhibit diminished pro-inflammatory cytokine response to LPS and induce robust proliferation of alloreactive T-cells. These proliferative alloreactive T-cells expressed greater levels of PD-1 and underwent increased programmed cell death as the concentration of BEN exposure increased. Alloreactive CD4+ T-cell death may be attributable to pre-cDC1s and provides a potential mechanism by which BEN+TBI conditioning limits GvHD and yields T-cells tolerant to host antigen.

Immunomodulatory Effects of Bendamustine in Hematopoietic Cell Transplantation.

Bendamustine (BEN) is a unique alkylating agent with efficacy against a broad range of hematological malignancies, although investigations have only recently started to delve into its immunomodulatory effects. These immunomodulatory properties of BEN in the context of hematopoietic cell transplantation (HCT) are reviewed here. Pre- and post-transplant use of BEN in multiple murine models have consistently resulted in reduced GvHD and enhanced GvL, with significant changes to key immunological cell populations, including T-cells, myeloid derived suppressor cells (MDSCs), and dendritic cells (DCs). Further, in vitro studies find that BEN enhances the suppressive function of MDSCs, skews DCs toward cDC1s, enhances Flt3 expression on DCs, increases B-cell production of IL-10, inhibits STAT3 activation, and suppresses proliferation of T- and B-cells. Overall, BEN has a broad range of immunomodulatory effects that, as they are further elucidated, may be exploited to improve clinical outcomes. As such, clinical trials are currently underway investigating new potential applications of BEN in the setting of allogeneic HCT.

Factors Affecting Site Use Preference of Grazing Cattle Studied from 2000 to 2020 through GPS Tracking: A Review.

Understanding the behaviour of grazing animals at pasture is crucial in order to develop management strategies that will increase the potential productivity of grazing systems and simultaneously decrease the negative impact on the environment. The objective of this review was to summarize and analyse the scientific literature that has addressed the site use preference of grazing cattle using global positioning systems (GPS) collars in the past 21 years (2000-2020) to aid the development of more sustainable grazing livestock systems. The 84 studies identified were undertaken in several regions of the world, in diverse production systems, under different climate conditions and with varied methodologies and animal types. This work presents the information in categories according to the main findings reviewed, covering management, external and animal factors driving animal movement patterns. The results showed that some variables, such as stocking rate, water and shade location, weather conditions and pasture (terrain and vegetation) characteristics, have a significant impact on the behaviour of grazing cattle. Other types of bio-loggers can be deployed in grazing ruminants to gain insights into their metabolism and its relationship with the landscape they utilise. Changing management practices based on these findings could improve the use of grasslands towards more sustainable and productive livestock systems.

Thermal limits for flight activity of field-collected Culicoides in the United Kingdom defined under laboratory conditions.

BACKGROUND: Culicoides biting midges (Diptera: Ceratopogonidae) are biological vectors of internationally important arboviruses and inflict biting nuisance on humans, companion animals and livestock. In temperate regions, transmission of arboviruses is limited by temperature thresholds, in both replication and dissemination of arboviruses within the vector and in the flight activity of adult Culicoides. This study aims to determine the cold-temperature thresholds for flight activity of Culicoides from the UK under laboratory conditions. METHODS: Over 18,000 Culicoides adults were collected from the field using 4 W down-draught miniature ultraviolet Centers for Disease Control traps. Populations of Culicoides were sampled at three different geographical locations within the UK during the summer months and again in the autumn at one geographical location. Activity at constant temperatures was assessed using a bioassay that detected movement of adult Culicoides towards an ultraviolet light source over a 24-h period. RESULTS: The proportion of active adult Culicoides increased with temperature but cold temperature thresholds for activity varied significantly according to collection season and location. Populations dominated by the subgenus Avaritia collected in South East England had a lower activity threshold temperature in the autumn (4 °C) compared with populations collected in the summer (10 °C). Within the subgenus Avaritia, Culicoides scoticus was significantly more active across all temperatures tested than Culicoides obsoletus within the experimental setup. Populations of Culicoides impunctatus collected in the North East of England were only active once temperatures reached 14 °C. Preliminary data suggested flight activity of the subgenus Avaritia does not differ between populations in South East England and those in the Scottish Borders. CONCLUSIONS: These findings demonstrate seasonal changes in temperature thresholds for flight and across different populations of Culicoides. These data, alongside that defining thresholds for virus replication within Culicoides, provide a primary tool for risk assessment of arbovirus transmission in temperate regions. In addition, the study also provides a comparison with thermal limits derived directly from light-suction trapping data, which is currently used as the main method to define adult Culicoides activity during surveillance.

IgA natural antibodies are produced following T-cell independent B-cell activation following stroke.

Up to 30% of stroke patients experience cognitive decline within one year of their stroke. There are currently no FDA-approved drugs that can prevent post-stroke cognitive decline, in part due to a poor understanding of the mechanisms involved. We have previously demonstrated that a B-lymphocyte response to stroke, marked by IgA + cells, can cause delayed cognitive dysfunction in mice and that a similar adaptive immune response occurs in the brains of some human stroke patients that suffer from vascular dementia. The stimuli which trigger B-lymphocyte activation following stroke, and their target antigens, are still unknown. Therefore, to learn more about the mechanisms by which B-lymphocytes become activated following stroke we first characterized the temporal kinetics of the B-lymphocyte, T-lymphocyte, and plasma cell (PC) response to stroke in the brain by immunohistochemistry (IHC). We discovered that B-lymphocyte, T-lymphocyte, and plasma cell infiltration within the infarct progressively increases between 2 and 7 weeks after stroke. We then compared the B-lymphocyte response to stroke in WT, MHCII-/-, CD4-/-, and MyD88-/- mice to determine if B-lymphocytes mature into IgA + PCs through a T-lymphocyte and MyD88 dependent mechanism. Our data from a combination of IHC and flow cytometry indicate that following stroke, a population of IgA + PCs develops independently of CD4 + helper T-lymphocytes and MyD88 signaling. Subsequent sequencing of immunoglobulin genes of individual IgA + PCs present within the infarct identified a novel population of natural antibodies with few somatic mutations in complementarity-determining regions. These findings indicate that a population of IgA + PCs develops in the infarct following stroke by B-lymphocytes interacting with one or more thymus independent type 2 (TI-2) antigens, and that they produce IgA natural antibodies.

Investigation of bovine ephemeral fever virus transmission by putative dipteran vectors under experimental conditions.

BACKGROUND: Bovine ephemeral fever virus (Rhabdoviridae: Ephemerovirus) (BEFV) causes bovine ephemeral fever (BEF), an economically important disease of cattle and water buffalo. Outbreaks of BEF in Africa, Australia, Asia and the Middle East are characterized by high rates of morbidity and highly efficient transmission between cattle hosts. Despite this, the vectors of BEFV remain poorly defined. METHODS: Colony lines of biting midges (Culicoides sonorensis) and mosquitoes (Aedes aegypti, Culex pipiens and Culex quinquefasciatus) were infected with a strain of BEFV originating from Israel by feeding on blood-virus suspensions and by intrathoracic inoculation. In addition, in vivo transmission of BEFV was also assessed by allowing C. sonorensis inoculated by the intrathoracic route to feed on male 6 month-old Holstein-Friesian calves. RESULTS: There was no evidence of BEFV replication within mosquitoes fed on blood/virus suspensions for mosquitoes of any species tested for each of the three colony lines. In 170 C. sonorensis fed on the blood/virus suspension, BEFV RNA was detected in the bodies of 13 individuals and in the heads of two individuals, indicative of fully disseminated infections and an oral susceptibility rate of 1.2%. BEFV RNA replication was further demonstrated in all C. sonorensis that were inoculated by the intrathoracic route with virus after 5, 6 or 7 days post-infection. Despite this, transmission of BEFV could not be demonstrated when infected C. sonorensis were allowed to feed on calves. CONCLUSIONS: No evidence for infection or dissemination of BEFV (bovine/Israel/2005-6) in mosquitoes of three different species was found. Evidence was found for infection of C. sonorensis by the oral route. However, attempts to transmit BEFV to calves from infected C. sonorensis failed. These results highlight the challenge of defining the natural vector of BEFV and of establishing an in vivo transmission model. The results are discussed with reference to the translation of laboratory-based studies to inference of vector competence in the field.

Bendamustine Conditioning Skews Murine Host DCs Toward Pre-cDC1s and Reduces GvHD Independently of Batf3.

Graft-versus-host disease (GvHD) remains the second leading cause of death in allogeneic hematopoietic stem cell transplantation recipients, highlighting the need for improved preventative strategies. Our laboratory has previously demonstrated in an experimental bone marrow transplantation (BMT) model that bendamustine combined with total body irradiation (BEN+TBI) is a safer alternative to cyclophosphamide with TBI (CY+TBI). The biological mechanisms of action of BEN have not been fully elucidated and likely involve multiple cell populations. Host dendritic cells (DCs) can prime naïve donor T-cells immediately following transplantation, making host DCs critical for the initiation phase of GvHD. We hypothesized that BEN+TBI conditioning favorably alters host DC composition to reduce GvHD. We demonstrate that host DCs treated with BEN+TBI induce less allogeneic T-cell proliferation than those conditioned with CY+TBI. We further show that BEN+TBI conditioning results in greater total numbers of all host DC subsets but with a more favorable composition compared to CY+TBI with significantly larger proportions of type 1 conventional DCs (cDC1), a highly regulatory DC subset capable of suppressing GvHD. Our studies using recipient Batf3 KO mice indicate that CD8α+ cDC1s are largely dispensable for the reduced GvHD following BEN+TBI conditioning. We found a higher frequency of host pre-cDC1s with BEN+TBI conditioning in both wild-type (WT) and Batf3 KO mice, which was inversely associated with GvHD. Additionally, we observed that BEN treatment results in greater expression of Flt3 receptor (CD135) on host DCs compared to CY, potentially contributing to the skewing of host DCs toward cDC1s. Further, BEN+TBI conditioning results in host cDCs with greater expression of PIR-B, an inhibitory receptor capable of preventing lethal GvHD. We conclude that BEN+TBI is a safer alternative to CY+TBI, resulting in a greater frequency of host pre-cDC1s and limiting GvHD.

Bendamustine with total body irradiation conditioning yields tolerant T-cells while preserving T-cell-dependent graft-versus-leukemia.

Graft-versus-host disease (GvHD) remains a significant impediment to allogeneic hematopoietic cell transplantation (HCT) success, necessitating studies focused on alleviating GvHD, while preserving the graft-versus-leukemia (GvL) effect. Based on our previous studies showing bendamustine with total body irradiation (BEN-TBI) conditioning reduces GvHD compared to the current clinical standard of care cyclophosphamide (CY)-TBI in a murine MHC-mismatched bone marrow transplantation (BMT) model, this study aimed to evaluate the role and fate of donor T-cells following BEN-TBI conditioning. We demonstrate that BEN-TBI reduces GvHD compared to CY-TBI independently of T regulatory cells (Tregs). BEN-TBI conditioned mice have a smaller proportion and less activated donor T-cells, with lower CD47 expression, early post-transplant, but no sustained phenotypic differences in T-cells. In BEN-TBI conditioned mice, donor T-cells gain tolerance specific to host MHC antigens. Though these T-cells are tolerant to host antigens, we demonstrate that BEN-TBI preserves a T-cell-dependent GvL effect. These findings indicate that BEN-TBI conditioning reduces GvHD without compromising GvL, warranting its further investigation as a potentially safer and more efficacious clinical alternative to CY-TBI.

Economic and Welfare Impacts of Providing Good Life Opportunities to Farm Animals.

Existing animal welfare standards for legislation and food certification programmes are primarily designed to avoid harms to the livestock, with minimal consideration given to their behavioural freedoms. Recent research has shown, however, that animal welfare should not only be evaluated by the absence of negative states but also by the presence of "good life" or positive experiences enjoyed by animals. The objective of the present study is to investigate the scientific validity and on-farm cost implications of utilising potential input-based measures of positive welfare as part of evaluation criteria for farm assurance schemes. Building upon the Farm Animal Welfare Council's concept of good life opportunities, an assessment was undertaken on 49 noncaged laying hen farms across the UK by measuring on-farm resources to facilitate positive experiences alongside commonly measured metrics for welfare outcomes. The financial cost of providing these resources on each enterprise was also estimated using a farm-scale costing tool. The results suggested that 63% of resource needs that facilitate the behaviour opportunities of laying hens are already being provided by these producers, far above legal and commercial requirements. This practice attracts no reward mechanism or direct financial benefit under the current market structure. Additional provision of opportunities was positively associated with behavioural outcomes, but only limited impact was observed on health and productivity measures. Economic modelling indicated that significant room exists to further improve welfare scores on these farms, on average by 97%, without incurring additional costs. Together we argue that these results can be seen as evidence of market failure since producers are providing positive welfare value to society that is not being currently recognised. It is therefore contended that measuring and rewarding the supply of good life opportunities could be a novel policy instrument to create an effective marketplace that appropriately recognises high welfare production.