Horse populations are severely underestimated in a region at risk of Hendra virus spillover.

OBJECTIVE: To identify the size and distribution of the horse population in the Northern Rivers Region of NSW, including changes from 2007 to 2021, to better understand populations at risk of Hendra virus transmission. METHODS: Census data from the 2007 Equine Influenza (EI) outbreak were compared with data collected annually by New South Wales Local Land Services (LLS) (2011-2021), and with field observations via road line transects (2021). RESULTS: The horse populations reported to LLS in 2011 (3000 horses; 0.77 horses/km2) was 145% larger than that reported during the EI outbreak in 2007 (1225 horses; 0.32 horses/km2). This was inconsistent with the 6% increase in horses recorded from 2011 to 2020 within the longitudinal LLS dataset. Linear modelling suggested the true horse population of this region in 2007 was at least double that reported at the time. Distance sampling in 2021 estimated the region's population at 10,185 horses (3.89 per km2; 95% CI = 4854-21,372). Field sampling and modelling identified higher horse densities in rural cropland, with the percentage of conservation land, modified grazing, and rural residential land identified as the best predictors of horse densities. CONCLUSIONS: Data from the 2007 EI outbreak no longer correlates to the current horse population in size or distribution and was likely not a true representation at the time. Current LLS data also likely underestimates horse populations. Ongoing efforts to further quantify and map horse populations in Australia are important for estimating and managing the risk of equine zoonoses.

The first patients treated with MR-CBCT soft-tissue matching in a MR-only prostate radiotherapy pathway.

INTRODUCTION: Magnetic Resonance (MR)-only radiotherapy for prostate cancer has previously been reported using fiducial markers for on-treatment verification. MR-Cone Beam Computed Tomography (CBCT) soft-tissue matching does not require invasive fiducial markers and enables MR-only treatments to other pelvic cancers. This study evaluated the first clinical implementation of MR-only prostate radiotherapy using MR-CBCT soft-tissue matching. METHODS: Twenty prostate patients were treated with MR-only radiotherapy using a synthetic (s)CT-optimised plan with MR-CBCT soft-tissue matching. Two MR sequences were acquired: small Field Of View (FOV) for target delineation and large FOV for organs at risk delineation, sCT generation and on-treatment verification. Patients also received a CT for validation. The prostate was independently contoured on the small FOV MR, copied to the registered CT and modified if there were MR-CT soft-tissue alignment differences (MR-CT volume). This was compared to the MR-only volume with a paired t-test. The treatment plan was recalculated on CT and the doses compared. Independent offline CT-CBCT matches for 5/20 fractions were performed by three therapeutic radiographers using the MR-only contours and compared to the online MR-CBCT matches using two one-sided paired t-tests for equivalence within ±1 mm. RESULTS: The MR-only volumes were significantly smaller than MR-CT (p = 0.003), with a volume ratio 0.92 ± 0.02 (mean ± standard error). The sCT isocentre dose difference to CT was 0.2 ± 0.1%. MR-CBCT soft-tissue matching was equivalent to CT-CBCT (p < 0.001), with differences of 0.1 ± 0.2 mm (vertical), -0.1 ± 0.2 mm (longitudinal) and 0.0 ± 0.1 mm (lateral). CONCLUSIONS: MR-only radiotherapy with soft-tissue matching has been successfully clinically implemented. It produced significantly smaller target volumes with high dosimetric and on-treatment matching accuracy. IMPLICATIONS FOR PRACTICE: MR-only prostate radiotherapy can be safely delivered without using invasive fiducial markers. This enables MR-only radiotherapy to be extended to other pelvic cancers where fiducial markers cannot be used.

Conditions affecting the timing and magnitude of Hendra virus shedding across pteropodid bat populations in Australia.

Understanding infection dynamics in animal hosts is fundamental to managing spillover and emergence of zoonotic infections. Hendra virus is endemic in Australian pteropodid bat populations and can be lethal to horses and humans. However, we know little about the factors driving Hendra virus prevalence in resevoir bat populations, making spillover difficult to predict. We use Hendra virus prevalence data collected from 13 000 pooled bat urine samples across space and time to determine if pulses of prevalence are periodic and synchronized across sites. We also test whether site-specific precipitation and temperature affect the amplitude of the largest annual prevalence pulses. We found little evidence for a periodic signal in Hendra virus prevalence. Although the largest amplitude pulses tended to occur over winter, pulses could also occur in other seasons. We found that Hendra virus prevalence was weakly synchronized across sites over short distances, suggesting that prevalence is driven by local-scale effects. Finally, we found that drier conditions in previous seasons and the abundance of Pteropus alecto were positively correlated with the peak annual values of Hendra virus prevalence. Our results suggest that in addition to seasonal effects, bat density and local climatic conditions interact to drive Hendra virus infection dynamics.

Persistent infections support maintenance of a coronavirus in a population of Australian bats (Myotis macropus).

Understanding viral transmission dynamics within populations of reservoir hosts can facilitate greater knowledge of the spillover of emerging infectious diseases. While bat-borne viruses are of concern to public health, investigations into their dynamics have been limited by a lack of longitudinal data from individual bats. Here, we examine capture-mark-recapture (CMR) data from a species of Australian bat (Myotis macropus) infected with a putative novel Alphacoronavirus within a Bayesian framework. Then, we developed epidemic models to estimate the effect of persistently infectious individuals (which shed viruses for extensive periods) on the probability of viral maintenance within the study population. We found that the CMR data analysis supported grouping of infectious bats into persistently and transiently infectious bats. Maintenance of coronavirus within the study population was more likely in an epidemic model that included both persistently and transiently infectious bats, compared with the epidemic model with non-grouping of bats. These findings, using rare CMR data from longitudinal samples of individual bats, increase our understanding of transmission dynamics of bat viral infectious diseases.

Ex vivo restimulation of human PBMC expands a CD3+CD4-CD8- γδ+ T cell population that can confound the evaluation of CD4 and CD8 T cell responses to vaccination.

The measurement of vaccine-induced humoral and CD4(+) and CD8(+) cellular immune responses represents an important correlate of vaccine efficacy. Accurate and reliable assays evaluating such responses are therefore critical during the clinical development phase of vaccines. T cells play a pivotal role both in coordinating the adaptive and innate immune responses and as effectors. During the assessment of cell-mediated immunity (CMI) in subjects participating in a large-scale influenza vaccine trial, we identified the expansion of an IFN-γ-producing CD3(+)CD4(-)CD8(-) γδ (+) T cell population in the peripheral blood of 90/610 (15%) healthy subjects. The appearance of CD3(+)CD4(-)CD8(-) γδ (+) T cells in the blood of subjects was transient and found to be independent of the study cohort, vaccine group, subject gender and ethnicity, and ex vivo restimulation conditions. Although the function of this population and relevance to vaccination are unclear, their inclusion in the total vaccine-specific T-cell response has the potential to confound data interpretation. It is thus recommended that when evaluating the induction of IFN-γ-producing CD4(+) and CD8(+) immune responses following vaccination, the CD3(+)CD4(-)CD8(-) γδ (+) T cells are either excluded or separately enumerated from the overall frequency determination.

Variation in radiotherapy target volume definition, dose to organs at risk and clinical target volumes using anatomic (computed tomography) versus combined anatomic and molecular imaging (positron emission tomography/computed tomography): intensity-modulated radiotherapy delivered using a tomotherapy Hi Art machine: final results of the VortigERN study.

AIMS: Contrast-enhanced computed tomography (CECT) is the current standard for delineating tumours of the head and neck for radiotherapy. Although metabolic imaging with positron emission tomography (PET) has been used in recent years, the studies were non-confirmatory in establishing its routine role in radiotherapy planning in the modern era. This study explored the difference in gross tumour volume and clinical target volume definitions for the primary and nodal volumes when FDG PET/CT was used as compared with CECT in oropharyngeal cancer cases. MATERIALS AND METHODS: Twenty patients with oropharyngeal cancers had a PET/CT scan in the treatment position after consent. Target volumes were defined on CECT scans by a consultant clinical oncologist who was blind to the PET scans. After obtaining inputs from a radiologist, another set of target volumes were outlined on the PET/CT data set. The gross and clinical target volumes as defined on the two data sets were then analysed. The hypothesis of more accurate target delineation, preventing geographical miss and comparative overlap volumes between CECT and PET/CT, was explored. The study also analysed the volumes of intersection and analysed whether there was any TNM stage migration when PET/CT was used as compared with CECT for planning. RESULTS: In 17 of 20 patients, the TNM stage was not altered when adding FDG PET information to CT. PET information prevented geographical miss in two patients and identified distant metastases in one case. PET/CT gross tumour volumes were smaller than CECT volumes (mean ± standard deviation: 25.16 cm(3) ± 35.8 versus 36.56 cm(3) ± 44.14; P < 0.015) for the primary tumour. Interestingly, our study showed no significant differences in gross tumour volume for T1/T2 disease, although differences in gross tumour volumes for advanced disease (T3/T4) were significant. The nodal target volumes (mean ± standard deviation: CECT versus PET/CT 32.48 cm(3) ± 36.63 versus 32.21 cm(3) ± 37.09; P > 0.86) were not statistically different. Similarity and discordance coefficients were calculated and are reported. CONCLUSION: PET/CT as compared with CECT could provide more clinically relevant information and prevent geographical miss when used for radiotherapy planning for advanced oropharyngeal tumours. Also, PET/CT provided a smaller better-defined target volume when compared with CECT. PET/CT-based volumes could therefore be used for treatment planning and targeted dose painting in oropharyngeal cancers.

Comparison of a simple dose-guided intervention technique for prostate radiotherapy with existing anatomical image guidance methods.

OBJECTIVES: A simple dose-guided intervention technique for prostate radiotherapy using an isodose overlay method combined with soft-tissue-based corrective couch shifts has been proposed previously. This planning study assesses the potential clinical impact of such a correction strategy. METHODS: 10 patients, each with 8-11 on-treatment CT studies (n=97), were assessed using this technique and compared with no intervention, bony anatomy intervention and soft-tissue intervention methods. Each assessment technique used a 4-mm action level for intervention. Outcomes were evaluated using measures of sensitivity, specificity and dosimetric effect, and compared across intervention techniques. Dosimetric effect was defined as the change in dosimetric coverage by the 95% isodose from the no intervention case of an evaluation construct called the verification target volume. RESULTS: Bony anatomy, soft tissue and dosimetric overlay-based interventions demonstrated sensitivity of 0.56, 0.73 and 1.00 and specificity of 0.64, 0.20 and 0.66, respectively. A detrimental dosimetric effect was shown in 7% of interventions for each technique, with benefit in 30%, 35% and 55% for bony anatomy, soft tissue and dosimetric overlay techniques, respectively. CONCLUSION: Used in conjunction with soft-tissue-based corrective couch shifts, the dosimetric overlay technique allows effective filtering out of dosimetrically unnecessary interventions, making it more likely that any intervention made will result in improved target volume coverage.

A dose distribution overlay technique for image guidance during prostate radiotherapy.

Adaptive radiotherapy involves altering the treatment plan according to variations in patient anatomy and set-up. This relies upon an accurate representation of the changing dose distribution within the patient, requiring a full dose recalculation. This work proposes a novel workflow using the planned dose distribution to assess dose coverage in three-dimensional verification CT studies acquired at the time of treatment delivery, using an overlay technique, in lieu of a recalculated dose distribution. The concept has been validated in a pilot study of 10 patients, each with 7-10 on-treatment CT studies. Differences between the geometric shape of the treatment plans for the 95% isodose and the 95% isodose obtained when the planned geometry was recalculated from the verification CT dataset were quantified. Dosimetric coverage of the verification clinical target volume (vCTV) was assessed for both the proposed overlay technique and the recalculated "delivered" dose distribution, and the conclusions on adequacy were compared. Results were consistent with geometric uncertainties of the dose calculation matrix (5 x 5 x 5 mm), suggesting that differences in the geometric shape of the 95% isodose are not significant for normal variations in patients' anatomy. Decisions on adequacy of vCTV coverage were consistent in 80 out of 87 cases, with discrepancies limited to a maximum of three axial slices per study within the range 0.5-4.5 mm (mean, 1.6 mm). The proposed dosimetric overlay technique has been validated and found to be an acceptable method of image-guided radiotherapy of the prostate suitable for effective implementation in the treatment clinic.

Dosimetric and treatment planning considerations for radiotherapy of the chest wall.

Radiotherapy treatment planning calculations of the chest wall are complex due to missing tissue, the thin chest wall and the presence of lung. The accuracy of calculated dose is dependent on the type of algorithm employed. This work evaluates a collapsed cone (CC) and a pencil beam (PB) convolution model for radiotherapy planning of the chest wall. Various irradiation geometries simulating the chest wall have been examined and calculations were compared with measurements with an ionization chamber in epoxy resin water substitute and in low-density lung substitute blocks. A retrospective treatment planning study comprising 6 patients was carried out to evaluate the differences in the dose distributions and monitor units predicted by the two algorithms. The calculated dose in unit density medium was within +/-1% for the CC model and up to +/-2% for PB. In low density medium and under full scatter conditions, CC overestimated the dose by 1% whereas PB overestimated the dose by 9%. In the tangential irradiation geometry with water and lung media, the PB overestimated dose to the isocentre by up to 10%, whereas the dose from CC was within 3%. From the treatment planning study calculated monitor units (MU) and doses were consistent with the experimental findings. The CC model is more accurate for radiotherapy treatment planning of the chest wall and especially when there is significant involvement of lung tissue.

Effect of B vitamins and genetics on success of in-vitro fertilisation: prospective cohort study.

BACKGROUND: There is a need to understand what affects the success of in-vitro fertilisation (IVF) and the rate of resulting twin births so that pregnancy rates can be improved and multiple gestations avoided. Our aim was to assess the role of B vitamins and genetics. METHODS: We did a prospective cohort study of 602 women undergoing fertility treatment. We assessed intake of folate and vitamin B12 with a questionnaire and measured their plasma and red-blood-cell concentrations by radioimmunoassay. We measured five B-vitamin-related gene variants in women who received treatment and in 932 women who conceived naturally. FINDINGS: The likelihood of a twin birth after IVF rose with increased concentrations of plasma folate (1.52, 1.01-2.28; p=0.032) and red-cell folate (1.28, 1.00-1.65; p=0.039). There was no association between folate and vitamin B12 levels and likelihood of a successful pregnancy. Women homozygous for the 1298 CC variant of methylenetetrahydro-folate reductase (MTHFR), rather than the AA variant, were less likely to produce a livebirth after IVF (0.24, 0.08-0.71; p=0.003) or to have had a previous pregnancy (0.42, 0.21-0.81; p=0.008). INTERPRETATION: Our findings suggest that MTHFR genotype is linked to a woman's potential to produce healthy embryos (possibly through interaction with genes related to DNA methylation). In women likely to have a successful IVF pregnancy, high folate status increases the likelihood of twin birth after multiple embryo transfer. Proposals to fortify the UK diet with folic acid could lead to an increase in the number of twins born after IVF.

Paediatric pelvic imaging: improvement in gonad shield placement by multidisciplinary audit.

BACKGROUND: In 1996, there were local reports of poor gonad protection for paediatric pelvic radiographs. OBJECTIVES: To investigate the nature of the problem and make necessary improvements. MATERIALS AND METHODS: A retrospective audit of 218 paediatric pelvic radiographs was undertaken in 1997. Each radiograph was assessed for the presence of a gonad shield, appropriateness of the device and its position. A multidisciplinary team was formed with representation from radiology, radiography, orthopaedics and medical physics to investigate ways of improving technique and reducing patient dose. These included radiographer training and the introduction of digital fluoroscopy as an alternative imaging technique in follow-up patients. There were further rounds of data collection in 1998 and 1999. RESULTS: In round 1, a gonad shield was present in 77.9 % of boys' films and 76 % of girls' films where one should have been, increasing to 85.2 % and 85.4 % respectively by round 3 of the audit (P < 0.05). Only 31.6 % of boys' devices and 21.9 % of girls' devices were correctly positioned in round 1, increasing to 78.3 % and 94.3 %, respectively, by round 3 of the audit (P < 0.05). After round 1, no inappropriate devices were used. CONCLUSION: Audit was an effective tool in gaining the resources needed to improve technique and reduce radiation exposure in children. The multidisciplinary approach was vital in the success of this project.

Paediatric pelvic imaging: optimisation of dose and technique using digital grid-controlled pulsed fluoroscopy.

BACKGROUND: An audit of paediatric pelvic radiographs identified deficiencies in gonad shield placement and radiographic technique. OBJECTIVE: A technique using grid-controlled fluoroscopy (GCF), with hard copy images in frame grab and digital spot image (DSI) format was evaluated to optimise gonad shield placement and reduce the dose given to children with Perthes disease and Developmental Hip Dysplasia (DDH) attending for pelvic radiography. MATERIALS AND METHODS: Phantom and patient dose surveys of conventional and fluoroscopic techniques were carried out. Image quality and radiation dose were compared for the frame grab and DSI techniques. Retrospective evaluation was undertaken to compare their clinical acceptability. RESULTS: Both fluoroscopic techniques gave considerably less radiation than conventional non-grid radiography (67-83%, P < 0.05). The frame grab technique gave less radiation than DSI (P < 0.05). There was no significant difference in the clinical acceptability scores of the DSI and frame grab images. CONCLUSION: Fluoroscopy acquired images are now used since the fluoroscopic techniques give much less dose than conventional radiography and provide images of sufficient quality for clinical assessment. Indeed, as there was no significant difference in clinical usefulness between the frame grab and DSI techniques, it is planned to use frame grab alone, thus gaining additional dose saving.

How should pathogen transmission be modelled?

Host-pathogen models are essential for designing strategies for managing disease threats to humans, wild animals and domestic animals. The behaviour of these models is greatly affected by the way in which transmission between infected and susceptible hosts is modelled. Since host-pathogen models were first developed at the beginning of the 20th century, the 'mass action' assumption has almost always been used for transmission. Recently, however, it has been suggested that mass action has often been modelled wrongly. Alternative models of transmission are beginning to appear, as are empirical tests of transmission dynamics.

The impact of cardiology on the collective effective dose in the North of England.

Two cardiology X-ray rooms were monitored with dose-area product meters as part of a Regional Patient Dosimetry Programme. Dose-area product measurements on over 2000 patients undergoing examinations in the cardiology rooms are presented. The data have been corrected according to patient size where possible. In room A mean dose-area product values for coronary angiography, coronary angioplasty, radiofrequency ablation and mitral valvuloplasty were found to be 47.7, 72.2, 91.1 and 161.9 Gy cm2 respectively. In room B mean dose-area product values for coronary angiography and coronary angioplasty were found to be 23.4 and 51.6 Gy cm2 respectively. Observational studies were used to deduce the typical projections and technique factors. This typical examination was used to simulate an angiogram from which it was possible to derive factors to convert measured dose-area product values into estimates of effective dose. In room A, the effective doses were estimated to be 9.4, 14.2, 17.3 and 29.3 mSv for coronary angiography, coronary angioplasty, radiofrequency ablation and mitral valvuloplasty, respectively. The effective doses during coronary angiography and coronary angioplasty, performed in room B, were found to be 4.6 and 10.2 mSv, respectively. A regional survey of the frequency of these cardiac procedures was performed. It was deduced that the annual collective effective dose from these cardiac procedures in the North of England, the former Northern Region, was 45.7 manSv.

Long-term culture of primary breast cancer in defined medium.

Over a period of 6 1/2 years between January 1986 and May 1992, 135 unselected primary breast cancers were cultured and of these 10 developed into cell lines. Six of the lines grew in defined serum-free medium, while the other four required supplementation with 0.5% fetal calf serum. Two of the lines are from the same breast, being derived from a local excision specimen and from a mastectomy specimen 12 months later. In addition, 12 lymph nodes containing metastatic breast cancer were cultured; one of these cultures became permanent in a defined serum-free medium. Oestrogen receptor (ER) status was negative in all but one of the tumours which grew successfully, and even in this case the derived cell line is ER negative. The epithelial nature of the lines has been confirmed by immunocytochemistry and by electron microscopy (EM), while their malignant nature is shown by morphology, unattached growth, chromosome analysis, and, in the case of the line from a lymph node metastasis, the absence of any benign source of epithelial cells.

Detecting disease and parasite threats to endangered species and ecosystems.

Ecologists have recently begun to acknowledge the importance of disease and parasites in the dynamics of populations. Diseases and parasites have probably been responsible for a number of extinctions on islands and on large land masses, but the problem has only been identified in retrospect. In contrast, endemic pathogens and parasites may operate as keystone species, playing a crucial role in maintaining the diversity of ecological communities and ecosystems. Will recent advances in the understanding of parasite population biology allow us to predict threats to endangered species and communities?