Shiga toxin sub-type 2a increases the efficiency of Escherichia coli O157 transmission between animals and restricts epithelial regeneration in bovine enteroids.

Specific Escherichia coli isolates lysogenised with prophages that express Shiga toxin (Stx) can be a threat to human health, with cattle being an important natural reservoir. In many countries the most severe pathology is associated with enterohaemorrhagic E. coli (EHEC) serogroups that express Stx subtype 2a. In the United Kingdom, phage type (PT) 21/28 O157 strains have emerged as the predominant cause of life-threatening EHEC infections and this phage type commonly encodes both Stx2a and Stx2c toxin types. PT21/28 is also epidemiologically linked to super-shedding (>103 cfu/g of faeces) which is significant for inter-animal transmission and human infection as demonstrated using modelling studies. We demonstrate that Stx2a is the main toxin produced by stx2a+/stx2c+ PT21/28 strains induced with mitomycin C and this is associated with more rapid induction of gene expression from the Stx2a-encoding prophage compared to that from the Stx2c-encoding prophage. Bacterial supernatants containing either Stx2a and/or Stx2c were demonstrated to restrict growth of bovine gastrointestinal organoids with no restriction when toxin production was not induced or prevented by mutation. Isogenic strains that differed in their capacity to produce Stx2a were selected for experimental oral colonisation of calves to assess the significance of Stx2a for both super-shedding and transmission between animals. Restoration of Stx2a expression in a PT21/28 background significantly increased animal-to-animal transmission and the number of sentinel animals that became super-shedders. We propose that while both Stx2a and Stx2c can restrict regeneration of the epithelium, it is the relatively rapid and higher levels of Stx2a induction, compared to Stx2c, that have contributed to the successful emergence of Stx2a+ E. coli isolates in cattle in the last 40 years. We propose a model in which Stx2a enhances E. coli O157 colonisation of in-contact animals by restricting regeneration and turnover of the colonised gastrointestinal epithelium.

Joint association between accelerometry-measured daily combination of time spent in physical activity, sedentary behaviour and sleep and all-cause mortality: a pooled analysis of six prospective cohorts using compositional analysis.

OBJECTIVE: To examine the joint associations of daily time spent in different intensities of physical activity, sedentary behaviour and sleep with all-cause mortality. METHODS: Federated pooled analysis of six prospective cohorts with device-measured time spent in different intensities of physical activity, sedentary behaviour and sleep following a standardised compositional Cox regression analysis. PARTICIPANTS: 130 239 people from general population samples of adults (average age 54 years) from the UK, USA and Sweden. MAIN OUTCOME: All-cause mortality (follow-up 4.3-14.5 years). RESULTS: Studies using wrist and hip accelerometer provided statistically different results (I2=92.2%, Q-test p<0.001). There was no association between duration of sleep and all-cause mortality, HR=0.96 (95% CI 0.67 to 1.12). The proportion of time spent in moderate to vigorous physical activity was significantly associated with lower risk of all-cause mortality (HR=0.63 (95% CI 0.55 to 0.71) wrist; HR=0.93 (95% CI 0.87 to 0.98) hip). A significant association for the ratio of time spent in light physical activity and sedentary time was only found in hip accelerometer-based studies (HR=0.5, 95% CI 0.42 to 0.62). In studies based on hip accelerometer, the association between moderate to vigorous physical activity and mortality was modified by the balance of time spent in light physical activity and sedentary time. CONCLUSION: This federated analysis shows a joint dose-response association between the daily balance of time spent in physical activity of different intensities and sedentary behaviour with all-cause mortality, while sleep duration does not appear to be significant. The strongest association is with time spent in moderate to vigorous physical activity, but it is modified by the balance of time spent in light physical activity relative to sedentary behaviour.

Integrating Sleep, Physical Activity, and Diet Quality to Estimate All-Cause Mortality Risk: A Combined Compositional Clustering and Survival Analysis of the National Health and Nutrition Examination Survey 2005-2006 Cycle.

We aimed to compare all-cause mortality risk across clusters of adults ≥50 years of age (n = 1,035) with common lifestyle behaviors patterns, enrolled in the US National Health and Nutrition Examination Survey (2005-2006). Log-ratio coordinates of 24-hour movement pattern and z scores of diet quality were used as input into a model-based clustering analysis. A Cox regression model was fitted to ascertain the all-cause mortality risk associated with each cluster. Participants were clustered into 4 groups: 1) a group characterized by a better physical activity profile and longer sleep duration coupled with an average diet quality (cluster 1); 2) a group with the poorest activity profile and shortest sleep but also the best diet quality (cluster 2); 3) another group featuring lower levels of activity of either intensity and higher levels of sedentary behavior and also a poor diet quality score (cluster 3); and 4) a group with an average diet quality and the best activity profile in the sample (cluster 4). A combination of a poorer diet and activity profile increased the prospective risk of all-cause mortality. Our findings emphasize the importance of considering the combination of diet quality and 24-hour movement patterns when developing interventions to reduce the risk of premature mortality.

Zero problems with compositional data of physical behaviors: a comparison of three zero replacement methods.

BACKGROUND: Researchers applying compositional data analysis to time-use data (e.g., time spent in physical behaviors) often face the problem of zeros, that is, recordings of zero time spent in any of the studied behaviors. Zeros hinder the application of compositional data analysis because the analysis is based on log-ratios. One way to overcome this challenge is to replace the zeros with sensible small values. The aim of this study was to compare the performance of three existing replacement methods used within physical behavior time-use epidemiology: simple replacement, multiplicative replacement, and log-ratio expectation-maximization (lrEM) algorithm. Moreover, we assessed the consequence of choosing replacement values higher than the lowest observed value for a given behavior. METHOD: Using a complete dataset based on accelerometer data from 1310 Danish adults as reference, multiple datasets were simulated across six scenarios of zeros (5-30% zeros in 5% increments). Moreover, four examples were produced based on real data, in which, 10 and 20% zeros were imposed and replaced using a replacement value of 0.5 min, 65% of the observation threshold, or an estimated value below the observation threshold. For the simulation study and the examples, the zeros were replaced using the three replacement methods and the degree of distortion introduced was assessed by comparison with the complete dataset. RESULTS: The lrEM method outperformed the other replacement methods as it had the smallest influence on the structure of relative variation of the datasets. Both the simple and multiplicative replacements introduced higher distortion, particularly in scenarios with more than 10% zeros; although the latter, like the lrEM, does preserve the ratios between behaviors with no zeros. The examples revealed that replacing zeros with a value higher than the observation threshold severely affected the structure of relative variation. CONCLUSIONS: Given our findings, we encourage the use of replacement methods that preserve the relative structure of physical behavior data, as achieved by the multiplicative and lrEM replacements, and to avoid simple replacement. Moreover, we do not recommend replacing zeros with values higher than the lowest observed value for a behavior.

A novel technique for retrospective genetic analysis of the response to vaccination or infection using cell-free DNA from archived sheep serum and plasma.

Genetic variation is associated with differences in disease resistance and susceptibility among individuals within a population. To date, molecular genetic analyses of host responses have relied on extraction of genomic DNA from whole blood or tissue samples. However, such samples are not routinely collected during large-scale field studies. We demonstrate that cell-free genomic DNA (cfDNA) may be extracted and amplified from archived plasma samples, allowing retrospective analysis of host genetic diversity. This technique was also applicable to archived serum samples up to 35 years old and to different ruminant species. As proof of concept, we used this cfDNA approach to genotype the major histocompatibility complex (MHC) class II DRB1 locus of 224 Merino sheep which had participated in field trials of a commercial Haemonchus contortus vaccine, Barbervax®, in Australia. This identified a total of 51 different DRB1 alleles and their relative frequencies. This is the first study to examine host MHC diversity using DNA extracted from archived plasma samples, an approach that may be applied to retrospective analyses of genetic diversity and responses to vaccination or infection across different species and populations.

Defining immune correlates during latent and active chlamydial infection in sheep.

Ovine enzootic abortion (OEA) caused by the obligate intracellular bacterial pathogen Chlamydia abortus (C. abortus), is an endemic disease in most sheep-rearing countries worldwide. Following infection, C. abortus establishes a complex host-pathogen interaction with a latent phase in non-pregnant sheep followed by an active disease phase in the placenta during pregnancy leading to OEA. Improved knowledge of the host-pathogen interactions at these different phases of disease will accelerate the development of new diagnostic tests and vaccines to control OEA. Current evidence indicates that cellular immunity is essential for controlling C. abortus infection. We have previously described a model of mucosal (intranasal) infection of non-pregnant sheep with C. abortus that replicates the latent and active phases of OEA. We have investigated antigen-specific recall responses of peripheral blood mononuclear cells (PBMC) in sheep infected with C. abortus via the intranasal route to determine how these change during the latent and active phases of disease. By analysing cytokines associated with the major CD4+ve Thelper (Th) cell subsets (Interferon-gamma (IFN-γ)/Th1; Interleukin (IL)-4/Th2; IL-17A/Th17; IL-10/Tregulatory), we show that there is selective activation of PBMC producing IFN-γ and/or IL-10 during the latent phase following infection. These cytokines are also elevated during the active disease phase and while they are produced by sheep that are protected from OEA, they are also produced by sheep that abort, highlighting the difficulties in finding specific cellular immunological correlates of protection for complex intracellular pathogens.

Mineral-nutrient relationships in African soils assessed using cluster analysis of X-ray powder diffraction patterns and compositional methods.

Soil mineral compositions are often complex and spatially diverse, with each mineral exhibiting characteristic chemical properties that determine the intrinsic total concentration of soil nutrients and their phyto-availability. Defining soil mineral-nutrient relationships is therefore important for understanding the inherent fertility of soils for sustainable nutrient management, and data-driven approaches such as cluster analysis allow for these relations to be assessed in new detail. Here the fuzzy-c-means clustering algorithm was applied to an X-ray powder diffraction (XRPD) dataset of 935 soils from sub-Saharan Africa, with each diffractogram representing a digital signature of a soil's mineralogy. Nine mineralogically distinct clusters were objectively selected from the soil mineralogy continuum by retaining samples exceeding the 75 % quantile of the membership coefficients in each cluster, yielding a dataset of 239 soils. As such, samples within each cluster represented mineralogically similar soils from different agro-ecological environments of sub-Saharan Africa. Mineral quantification based on the mean diffractogram of each cluster illustrated substantial mineralogical diversity between the nine groups with respect to quartz, K-feldspar, plagioclase, Fe/Al/Ti-(hydr)oxides, phyllosilicates (1:1 and 2:1), ferromagnesians, and calcite. Mineral-nutrient relationships were defined using the clustered XRPD patterns and corresponding measurements of total and/or extractable (Mehlich-3) nutrient concentrations (B, Mg, K, Ca, Mn, Fe, Ni, Cu and Zn) in combination with log-ratio compositional data analysis. Fe/Al/Ti/Mn-(hydr)oxides and feldspars were found to be the primary control of total nutrient concentrations, whereas 2:1 phyllosilicates were the main source of all extractable nutrients except for Fe and Zn. Kaolin minerals were the most abundant phyllosilicate group within the dataset but did not represent a nutrient source, which reflects the lack of nutrients within their chemical composition and their low cation exchange capacity. Results highlight how the mineral composition controls the total nutrient reserves and their phyto-availability in soils of sub-Saharan Africa. The typical characterisation of soils and their parent material based on the clay particle size fraction (i.e. texture) and/or the overall silica component (i.e. acid and basic rock types) alone may therefore mask the intricacies of mineral contributions to soil nutrient concentrations.

MALDIrppa: quality control and robust analysis for mass spectrometry data.

Summary: This R package helps to implement a robust approach to deal with mass spectrometry (MS) data. It is aimed at alleviating reproducibility issues and pernicious effects of deviating signals on both data pre-processing and downstream data analysis. Based on robust statistical methods, it facilitates the identification and filtering of low-quality mass spectra and atypical peak profiles as well as monitoring and data handling through pre-processing, which extends existing computational tools for high-throughput data. Availability and implementation: MALDIrppa is implemented as a package for the R environment for data analysis and it is freely available to download from the CRAN repository at https://CRAN.R-project.org/package=MALDIrppa. Contact: javier.palarea@bioss.ac.uk.

Enhancing the toolbox to study IL-17A in cattle and sheep.

The development of methods to detect cytokine expression by T cell subsets in ruminants is fundamental to strategic development of new livestock vaccines for prevention of infectious diseases. It has been possible to detect T cell expression of IFN-γ, IL-4 and IL-10 in ruminants for many years but methods to detect expression of IL-17A are relatively limited. To address this gap in capability we have cloned bovine and ovine IL-17A cDNAs and expressed biologically-active recombinant proteins in Chinese Hamster Ovary (CHO) cells. We used the transfected CHO cells to screen commercially-available antibodies for their ability to detect IL-17A expression intracellularly and in culture supernates. We demonstrate that an ELISA for bovine IL-17A detects native ovine IL-17A. Moreover, the constituent polyclonal antibodies (pabs) in the ELISA were used to enumerate peripheral blood mononuclear cells (PBMC) expressing IL-17A from cattle and sheep by ELISpot. We identified two monoclonal antibodies (mabs) that detect recombinant intracellular IL-17A in CHO cells by flow cytometry. One of these mabs was used to detect native intracellular IL-17A expression in PBMC in conjunction with cell surface phenotyping mabs [CD4+ve, CD8+ve and Workshop Cluster 1 (WC-1)+ve gamma-delta (γδ)] we show that distinct T cell subsets in cattle (defined as CD4+ve, CD8+ve or WC-1+ve) and sheep (defined as CD4+ve or WC-1+ve) can express IL-17A following activation. These novel techniques provide a solid basis to investigate IL-17A expression and define specific CD4+ve T cell subset activation in ruminants.

Inflammatory infiltration into placentas of Neospora caninum challenged cattle correlates with clinical outcome of pregnancy.

Infection with Neospora caninum stimulates host cell-mediated immune responses, which may be responsible for placental damage leading to bovine abortion. The aim of this study was to compare immune responses in the bovine placenta, following experimental infection in different stages of pregnancy. Placentomes were examined by immunohistochemistry and inflammation in early gestation was generally moderate to severe, particularly in the placentas carrying non-viable foetuses, whereas it was milder in later stages, mainly characterised by the presence of CD3+, CD4+ and γδ T-cells. This distinctive cellular immune response may explain the milder clinical outcome observed when animals are infected in later gestation.

A curated multivariate approach to study efficacy and optimisation of a prototype vaccine against teladorsagiasis in sheep.

This work discusses and demonstrates the novel use of multivariate analysis and data dimensionality reduction techniques to handle the variety and complexity of data generated in efficacy trials for the development of a prototype vaccine to protect sheep against the Teladorsagia circumcincta nematode. A curated collection of data dimension reduction and visualisation techniques, in conjunction with sensible statistical modelling and testing which explicitly model key features of the data, offers a synthetic view of the relationships between the multiple biological parameters measured. New biological insight is gained into the patterns and associations involving antigen-specific antibody levels, antibody avidity and parasitological parameters of efficacy that is not achievable by standard statistical practice in the field. This approach can therefore be used to guide vaccine refinement and simplification through identifying the most immunologically relevant antigens, and it can be analogously implemented for similar studies in other areas. To facilitate this, the associated data and computer codes written for the R open system for statistical computing are made freely available.

Association between musculoskeletal pain and exposures to awkward postures during work: a compositional analysis approach.

OBJECTIVES: This study aimed to explore the association between arm elevation and neck/shoulder pain, and trunk forwarding bending and low back pain among home care workers. METHODS: Home care workers (N = 116) from 11 home care units in Trondheim, Norway, filled in pain assessment and working hours questionnaire, and wore 3 accelerometers for up to 7 consecutive days. Work time was partitioned into upright awkward posture, nonawkward posture, and nonupright time, i.e. sitting. Within a compositional approach framework, posture time compositions were expressed in terms of log-ratio coordinates for statistical analysis and modeling. Poisson generalized linear mixed models were used to analyze the relationship between arm elevation in upright postures and neck/shoulder pain, and between trunk forward bending in upright postures and low back pain, respectively. Isotemporal substitution analysis was used to investigate the association of pain assessment with the reallocation of time spent in the different postures. RESULTS: Time spent in awkward postures was modest, especially for the more extreme angles (60° and 90°). Adjusting for age, gender, and body mass index, our study suggested that the compositions of time spent by home care workers in awkward postures were significantly associated with pain assessment (P < 0.01). Isotemporal substitution analysis showed that reallocating 5 min from upright posture with arms elevated below to above 60° and 90° was associated with a 6.8% and 19.9% increase in the neck/shoulder pain score, respectively. Reallocating 5 min from a forward bending posture while upright below to above 30°, 60°, and 90° was associated with 1.8%, 3.5%, and 4.0% increase in low back pain, respectively. CONCLUSIONS: Although the exposure to awkward postures was modest, our results showed an association between increased time spent in awkward postures and an increase in neck/shoulder pain and low back pain in home care workers. As musculoskeletal pain is the leading cause of sickness absence, these findings suggest that home care units could benefit from re-organizing work to avoid excessive arm elevation and trunk forward bending in workers.

Evaluation of species-specific polyclonal antibodies to detect and differentiate between Neospora caninum and Toxoplasma gondii.

Neosporosis and toxoplasmosis are major causes of abortion in livestock worldwide, leading to substantial economic losses. Detection tools are fundamental to the diagnosis and management of those diseases. Current immunohistochemistry (IHC) tests, using sera raised against whole parasite lysates, have not been able to distinguish between Toxoplasma gondii and Neospora caninum. We used T. gondii and N. caninum recombinant proteins, expressed in Escherichia coli and purified using insoluble conditions, to produce specific polyclonal rabbit antisera. We aimed to develop species-specific sera that could be used in IHC on formalin-fixed, paraffin-embedded (FFPE) tissue sections to improve the diagnosis of ruminant abortions caused by protozoa. Two polyclonal rabbit sera, raised against recombinant proteins, anti-Neospora-rNcSRS2 and anti-Toxoplasma-rTgSRS2, had specificity for the parasite they were raised against. We tested the specificity for each polyclonal serum using FFPE tissue sections known to be infected with T. gondii and N. caninum. The anti-Neospora-rNcSRS2 serum labeled specifically only N. caninum-infected tissue blocks, and the anti-Toxoplasma-rTgSRS2 serum was specific to only T. gondii-infected tissues. Moreover, tissues from 52 cattle and 19 sheep previously diagnosed by lesion profiles were tested using IHC with our polyclonal sera and PCR. The overall agreement between IHC and PCR was 90.1% for both polyclonal anti-rNcSRS2 and anti-rTgSRS2 sera. The polyclonal antisera were specific and allowed visual confirmation of protozoan parasites by IHC, but they were not as sensitive as PCR testing.

Phenotypic characterisation of the cellular immune infiltrate in placentas of cattle following experimental inoculation with Neospora caninum in late gestation.

Despite Neospora caninum being a major cause of bovine abortion worldwide, its pathogenesis is not completely understood. Neospora infection stimulates host cell-mediated immune responses, which may be responsible for the placental damage leading to abortion. The aim of the current study was to characterize the placental immune response following an experimental inoculation of pregnant cattle with N. caninum tachyzoites at day 210 of gestation. Cows were culled at 14, 28, 42 and 56 days post inoculation (dpi). Placentomes were examined by immunohistochemistry using antibodies against macrophages, T-cell subsets (CD4, CD8 and γδ), NK cells and B cells. Macrophages were detected mainly at 14 days post inoculation. Inflammation was generally mild and mainly characterized by CD3+, CD4+ and γδ T-cells; whereas CD8+ and NK cells were less numerous. The immune cell repertoire observed in this study was similar to those seen in pregnant cattle challenged with N. caninum at early gestation. However, cellular infiltrates were less severe than those seen during first trimester Neospora infections. This may explain the milder clinical outcome observed when animals are infected late in gestation.

Compositional functional regression and isotemporal substitution analysis: Methods and application in time-use epidemiology.

The distribution of time that people spend in physical activity of various intensities has important health implications. Physical activity (commonly categorised by the intensity into light, moderate and vigorous physical activity), sedentary behaviour and sleep, should not be analysed separately, because they are parts of a time-use composition with a natural constraint of 24 h/day. To find out how are relative reallocations of time between physical activity of various intensities associated with health, herewith we describe compositional scalar-on-function regression and a newly developed compositional functional isotemporal substitution analysis. Physical activity intensity data can be considered as probability density functions, which better reflects the continuous character of their measurement using accelerometers. These probability density functions are characterised by specific properties, such as scale invariance and relative scale, and they are geometrically represented using Bayes spaces with the Hilbert space structure. This makes possible to process them using standard methods of functional data analysis in the L2 space, via centred logratio (clr) transformation. The scalar-on-function regression with clr transformation of the explanatory probability density functions and compositional functional isotemporal substitution analysis were applied to a dataset from a cross-sectional study on adiposity conducted among school-aged children in the Czech Republic. Theoretical reallocations of time to physical activity of higher intensities were found to be associated with larger and more progressive expected decreases in adiposity. We obtained a detailed insight into the dose-response relationship between physical activity intensity and adiposity, which was enabled by using the compositional functional approach.

Measuring behaviour in hens using an ethogram to assess analgesia during further refinement of a high welfare, on-hen, poultry red mite feeding device.

Background: To refine an on-hen mite feeding device, an ethogram was employed to measure the reactions of hens during a routine experimental procedure (feather plucking) and to assess effects of analgesic cream on those reactions. Methods: Three experimental groups were used; one treated with EMLA 5% before plucking ("EMLA group"); one with aqueous cream ("placebo group") and a "no treatment" group. Behaviours were measured and compared on three days: 'dummy handling day' i.e. no plucking; 'plucking day', plucking the left thigh; and 'treatment day' i.e with right thighs plucked post-treatment. Poultry red mite feeding assays were performed to examine effect of creams on mite feeding rates, mortality and fecundity. All data were analysed using generalised linear (mixed) modelling approaches. Results: Use of the ethogram demonstrated no significant difference in hen behaviours in the EMLA group between dummy handling day and treatment day (p = 0.949) alongside a significant reduction in measured behaviours between plucking day and treatment day in the same group (p = 0.028). There was a statistically significant increase in measured behaviours from the dummy handling day to the plucking day in both placebo (p = 0.011) and no treatment group (p < 0.001). Effect sizes and directions were similar between dummy handling and treatment days in the 'placebo' and 'no treatment' groups, though not statistically significant (placebo, p = 0.064; no treatment p = 0.069). Mite feeding in the EMLA group was significantly lower than in the no treatment group in feeding assay 1 (p = 0.029) only. Mite mortality and fertility were unaffected. Conclusions: The ethogram successfully measured changes in observed behaviours between the dummy handling session and procedures. No adverse effects of EMLA cream on hens were demonstrated at 3mg/kg in hens. Use of analgesia for this routine procedure improves hens' experiences during experimental trials.

Blue prescription: A pilot study of health benefits for oncological patients of a short program of activities involving the sea.

Performing outdoor activities in blue spaces can help improve human health and mental well-being by reducing stress and promoting social relationships. The number of people surviving cancer has increased globally to experience this disease as a life-changing and chronic condition with physical and psychosocial symptoms that have negative impacts on their quality of life. While there has been a growth of programs in green spaces to meet the needs of cancer patients, such as follow-up post-treatment care, support groups and physical activity programs, very few studies have examined the effects of activities involving the sea for the health and well-being of oncology patients. This is the first study to evaluate whether different outdoor activities in blue spaces can benefit oncological patients' physical and mental health using smartwatches, sphygmomanometers and Profile of Mood States (POMS) questionnaires. We assessed changes in blood pressure, heart rate, sleep quality and mental health of 16 patients after twelve sessions of three different activities (walking, beach and snorkelling) and four sessions of a control activity. While no significant differences between activities were observed in terms of the data gathered by the smartwatches, a gradient of positive results for human mental health was observed towards exposure to a blue space, assessed through POMS questionnaires. Results show that exposure to blue spaces contributes to tension and anger reduction and improves the vigour mood state of oncology patients. No significant increases in patients' heart rate were recorded after the beach and snorkelling activities, with results similar to the control activity, suggesting that the contribution may be to participants' relaxation.

Efficacy of Phase I and Phase II Coxiella burnetii Bacterin Vaccines in a Pregnant Ewe Challenge Model.

The bacterium Coxiella burnetii can cause the disease Q-fever in a wide range of animal hosts. Ruminants, including sheep, are thought to play a pivotal role in the transmission of C. burnetii to humans; however, the only existing livestock vaccine, namely, Coxevac® (Ceva Animal Health Ltd., Libourne, France), a killed bacterin vaccine based on phase I C. burnetii strain Nine-Mile, is only approved for use in goats and cattle. In this study, a pregnant ewe challenge model was used to determine the protective effects of Coxevac® and an experimental bacterin vaccine based on phase II C. burnetii against C. burnetii challenge. Prior to mating, ewes (n = 20 per group) were vaccinated subcutaneously with either Coxevac®, the phase II vaccine, or were unvaccinated. A subset of pregnant ewes (n = 6) from each group was then challenged 151 days later (~100 days of gestation) with 106 infectious mouse doses of C. burnetii, Nine-Mile strain RSA493. Both vaccines provided protection against C. burnetii challenge as measured by reductions in bacterial shedding in faeces, milk and vaginal mucus, and reduced abnormal pregnancies, compared to unvaccinated controls. This work highlights that the phase I vaccine Coxevac® can protect ewes against C. burnetii infection. Furthermore, the phase II vaccine provided comparable levels of protection and may offer a safer and cost-effective alternative to the currently licensed vaccine.

High rate of transplacental infection and transmission of Neospora caninum following experimental challenge of cattle at day 210 of gestation.

In order to investigate the pathogenesis of neosporosis following a primary infection in late pregnancy, cattle were subcutaneously challenged with 5 × 108Neospora caninum (NC1 isolate) tachyzoites at day 210 of gestation and serial necropsies were then carried out at 14, 28, 42 and 56 days post-infection (dpi). No abortions occurred and all the foetuses were viable at the time of euthanasia. There was a high rate of vertical transmission, as parasites were detected by immunohistochemical labelling and PCR in all the foetuses from 28 dpi. Focal necrotic lesions were observed in the placentomes of the placenta from 28 dpi and showed resolution during later time points, denoted by infiltration of inflammatory cells at 42 dpi and fibrosis at 56 dpi. Foetuses at 28 and 42 dpi showed scarce and isolated lesions which are unlikely to represent a threat to foetal viability. No lesions were observed in the foetuses at 14 or 56 dpi suggesting control of the infection and resolution of the lesions by maternal and foetal immune responses. Once infection was established, it could not be cleared from the host and vertical transmission of the parasite occurred in all infected hosts. Parasite was detected in the placenta at 28 dpi, while in previous experimental infections of cattle at day 70 and 140 of gestation using the same challenge model, it was already present at day 14 post infection. This suggests that a change in the maternal immune response plays a crucial role in limiting the initial infection during the last term of pregnancy.

Compositional analysis of the association between mortality and 24-hour movement behaviour from NHANES.

AIMS: Previous prospective studies of the association between mortality and physical activity have generally not fully accounted for the interplay between movement behaviours. A compositional data modelling approach accounts for relative scale and co-dependency in time-use data across physical activity behaviours of the 24-hour day. METHODS: A prospective analysis of the National Health and Nutrition Examination Survey 2005-2006 on N = 1468 adults (d = 135 deaths) in ages 50-79 years was undertaken using compositional Cox regression analysis. Daily time spent in sedentary behaviour, light intensity (LIPA) and moderate-to-vigorous physical activity (MVPA) was determined from waist-mounted accelerometer data (Actigraph 7164) and supplemented with self-reported sleep data to determine the daily time-use composition. RESULTS: The composition of time spent in sedentary behaviour, LIPA, MVPA and sleep was associated with mortality rate after allowing for age and sex effects (p < 0.001), and remained significant when other lifestyle factors were added (p < 0.001). This was driven primarily by the preponderance of MVPA; however, significant changes are attributable to LIPA relative to sedentary behaviour and sleep, and sedentary behaviour relative to sleep. The final ratio ceased to be statistically significant after incorporating lifestyle factors. The preponderance of MVPA ceased to be statistically significant after incorporating health at outset and physical limitations on movement. CONCLUSIONS: An association is inferred between survival rate and the physical activity composition of the day. The MVPA time share is important, but time spent in LIPA relative to sedentary behaviour and sleep is also a significant factor. Increased preponderance of MVPA may have detrimental associations at higher levels of MVPA.