Detection of male genital schistosomiasis (MGS) associated with human, zoonotic and hybrid schistosomes in Southern Malawi.

BACKGROUND: Male Genital Schistosomiasis (MGS) remains an often-overlooked chronic sequela of urogenital schistosomiasis in endemic areas of sub-Saharan Africa. As part of a 2-year longitudinal study on Hybridization of UroGenital Schistosomiasis (HUGS) in Malawi, a MGS sub-study was conducted to assess whether hybrid schistosomes were incriminated. METHODS: During recruitment, demographic, health and socio-economic data were collected through individual questionnaire interviews in Mthawira community from Nsanje District along Shire River and Samama community from Mangochi District along Lake Malawi shoreline. Urine and semen samples were collected and analysed to determine the identity of schistosome infection. Urine filtration and microscopy, direct microscopy of semen and its sediments (after centrifugation) were performed. Thereafter, the sediments were examined by molecular DNA analysis with a novel two-tube real-time PCR assay. The participants were also screened for Human papilloma virus (HPV) and other sexually transmitted infections (STIs). RESULTS: Twenty-two men were recruited for the sub-study, 8 in Nsanje District and 14 in Mangochi District, with a median age of 22.0 years. By microscopy, ten (45.7%) participants had Schistosoma ova in their urine, 11 (50.0%) in semen while 16 (72.7%) were positive by real-time PCR. One participant had both S. haematobium and S. mattheei ova in his semen, three showed symptoms, and one had a mixed infection of S. mansoni and possible S. haematobium-S. mattheei hybrid. Twelve men had detectable high-risk HPV serotypes 16, 18 and others while six had Trichomonas vaginalis and other STIs. CONCLUSION: Zoonotic and hybrid schistosomes can cause MGS similar to human schistosomes, which can be co-infected with HPV and STIs, thereby posing a new challenge in diagnosis, management and control measures in resource poor settings. Increased awareness of these infections among local communities and primary healthcare workers and improvement of disease management are needed and advocated.

Revealing bovine schistosomiasis in Malawi: Connecting human and hybrid schistosomes within cattle.

In Malawi, the putative origin of a newly described Schistosoma haematobium-mattheei hybrid human schistosome was assessed upon a seminal molecular parasitological survey of cattle. Using miracidia hatch test (MHT) and carcass inspection at slaughter, mean prevalence of bovine schistosomiasis was 49.1% (95% CI: 43.7-54.6%) and 10.3% (95% CI: 6.0-16.2%) respectively, though significant spatial heterogeneity was noted. Approximately 2.0% of infected cattle, and only those from Mangochi District, shed S. haematobium-mattheei and/or S. haematobium in faeces. To quantify schistosome (re)infection dynamics, where a S. haematobium-mattheei hybrid was present, we undertook a novel pilot GPS-datalogging sub-study within a specific herd of cattle (n = 8) on the Lake Malawi shoreline, alongside a praziquantel (40 mg/kg) treatment efficacy spot check. At sub-study baseline, all GPS-tagged cattle had proven daily water contact with the lake. Each animal was patently infected upon MHT, with older animals shedding less miracidia. At one month review, whilst parasitological cure was 100.0%, from six weeks onwards, (re)infection was first noted in the youngest animal. By three-month review, all animals were patently (re)infected though only miracidia of S. mattheei were recovered, albeit in much lower numbers. To conclude, infection with S. mattheei is particularly common in cattle and demonstrates a previously cryptic burden of bovine schistosomiasis. Within Mangochi District, bovine transmission of both S. haematobium-mattheei hybrids and S. haematobium are now incriminated, with unequivocal evidence of contemporary zoonotic spill-over. Future control of urogenital schistosomiasis here in the southern region needs to develop, then successfully integrate, a One Health approach with appropriate mitigating strategies to reduce and/or contain bovine schistosomiasis transmission.

The importance of place-kicking in Women's International Rugby Union.

Despite the growing popularity of women's rugby, there is a lack of research understanding the contribution of place-kicking to match outcomes. This study aims to establish the characteristics and contribution of place-kicking to women's international Rugby Union and evaluate the performance of place-kickers while accounting for factors that contribute to kick difficulty. Data from 674 place-kicks across 80 matches were analysed. A binomial generalised linear mixed model (GLMM) was used to predict the probability of kick success. 60.5% of place-kicks were successful, and they contributed 23.9% of all points scored; conversions accounted for 16.8% and penalties 7.1%. Kick success percentages for conversions (56.9%) and penalties (78.3%) significantly differed (p < 0.01). Kick distance and angle were significant (p < 0.01) predictors of kick success and the GLMM had a prediction accuracy of 73.6%. The performance rankings of kickers changed when comparing observed and expected success, highlighting the need to consider contextual factors contributing to kick difficulty when evaluating performance. The GLMM results provide valuable insights for coaches and players to make informed decisions, for example, whether to attempt a place-kick when a penalty is awarded, by enabling predictions of place-kick success. This could enhance a team's chances of winning matches.

A geospatial analysis of local intermediate snail host distributions provides insight into schistosomiasis risk within under-sampled areas of southern Lake Malawi.

BACKGROUND: Along the southern shoreline of Lake Malawi, the incidence of schistosomiasis is increasing with snails of the genera Bulinus and Biomphalaria transmitting urogenital and intestinal schistosomiasis, respectively. Since the underlying distribution of snails is partially known, often being focal, developing pragmatic spatial models that interpolate snail information across under-sampled regions is required to understand and assess current and future risk of schistosomiasis. METHODS: A secondary geospatial analysis of recently collected malacological and environmental survey data was undertaken. Using a Bayesian Poisson latent Gaussian process model, abundance data were fitted for Bulinus and Biomphalaria. Interpolating the abundance of snails along the shoreline (given their relative distance along the shoreline) was achieved by smoothing, using extracted environmental rainfall, land surface temperature (LST), evapotranspiration, normalised difference vegetation index (NDVI) and soil type covariate data for all predicted locations. Our adopted model used a combination of two-dimensional (2D) and one dimensional (1D) mapping. RESULTS: A significant association between normalised difference vegetation index (NDVI) and abundance of Bulinus spp. was detected (log risk ratio - 0.83, 95% CrI - 1.57, - 0.09). A qualitatively similar association was found between NDVI and Biomphalaria sp. but was not statistically significant (log risk ratio - 1.42, 95% CrI - 3.09, 0.10). Analyses of all other environmental data were considered non-significant. CONCLUSIONS: The spatial range in which interpolation of snail distributions is possible appears < 10km owing to fine-scale biotic and abiotic heterogeneities. The forthcoming challenge is to refine geospatial sampling frameworks with future opportunities to map schistosomiasis within actual or predicted snail distributions. In so doing, this would better reveal local environmental transmission possibilities.

Development, validation, and pilot application of a high throughput molecular xenomonitoring assay to detect Schistosoma mansoni and other trematode species within Biomphalaria freshwater snail hosts.

Schistosomiasis is a neglected tropical disease (NTD) caused by infection with parasitic trematodes of the genus Schistosoma that can lead to debilitating morbidity and mortality. The World Health Organization recommend molecular xenomonitoring of Biomphalaria spp. freshwater snail intermediate hosts of Schistosoma mansoni to identify highly focal intestinal schistosomiasis transmission sites and monitor disease transmission, particularly in low-endemicity areas. A standardised protocol to do this, however, is needed. Here, two previously published primer sets were selected to develop and validate a multiplex molecular xenomonitoring end-point PCR assay capable of detecting S. mansoni infections within individual Biomphalaria spp. missed by cercarial shedding. The assay proved highly sensitive and highly specific in detecting and amplifying S. mansoni DNA and also proved highly sensitive in detecting and amplifying non-S. mansoni trematode DNA. The optimised assay was then used to screen Biomphalaria spp. collected from a S. mansoni-endemic area for infection and successfully detected S. mansoni infections missed by cercarial shedding as well as infections with non-S. mansoni trematodes. The continued development and use of molecular xenomonitoring assays such as this will aid in improving disease control efforts, significantly reducing disease-related morbidities experienced by those in schistosomiasis-endemic areas.

Human cercarial dermatitis (HCD) in the UK: an overlooked and under-reported nuisance?

BACKGROUND: Human cercarial dermatitis (HCD) is a clinical disease typically caused by skin-penetrative larvae of avian schistosomes. Its geographical epidemiology is firmly tied with that of infected freshwater intermediate snail hosts. To better understand the current distribution of HCD and its level of nuisance in the UK, we undertook a systematic literature review. METHODS: Following PRIMSA guidelines, PubMed and Scopus databases were searched with keywords "human cercarial dermatitis" OR "swimmer's itch" AND "United Kingdom". Articles about imported cases of HCD, or HCD outside the UK, were not formally included. RESULTS: A total of 30 articles were initially identified. A further two were gained by inspection of all citations. After screening, eight publications were analysed where the location, number of cases and putative avian schistosome species incriminated were tabulated. HCD is mainly found in the south of England, though gaps in evidence and reporting remain across the UK. CONCLUSIONS: Despite its noted recent rise in open water swimmers, published literature on HCD across the UK is sparse; this condition is both overlooked and under-reported. We therefore recommend establishing a national database that raises awareness and encourages self-reporting of this nuisance disease.

Post-traumatic stress disorder is more likely in younger patients undergoing lower limb arthroplasty: impact on function and quality of life.

INTRODUCTION: The aim of this study was to assess whether PTSD was associated with preoperative and/or postoperative joint-specific function and health-related quality of life (HRQoL) in patients undergoing total hip arthroplasty (THA) and total knee arthroplasty (TKA) and whether there were associated preoperative factors. METHODS: A retrospective study was conducted at a single centre using an established arthroplasty database over a 2-year period. Patients undergoing THA and TKA completed pre and 1-year postoperative Oxford hip/knee scores and EuroQoL questionnaire (EQ-5D) to assess joint specific function and HRQoL. Postoperatively, patients completed the self-reported PTSD Checklist for DSM-5 (PCL-5) questionnaire where a score of 31 or greater was used to determine a provisional diagnosis of PTSD. RESULTS: There were 1244 THA and 1356 TKA patients, of which 42 (3.4%) and 54 (4.0%) had a PCL-5 score of ≥ 31, respectively (PTSD groups). Younger age was associated (p < 0.001) with PTSD for both THA (mean difference (MD) 9.9, 95%CI 6.7-13.0) and TKA (MD 4.6, 95%CI 2.2-6.9), which remained significant when adjusting for confounding variables (THA: p < 0.001; TKA: p = 0.020). The preoperative Oxford (THA:MD 4.9, p < 0.001; TKA:MD 5.7, p < 0.001) and EQ-5D scores (THA:MD 0.378, p < 0.001; TKA:MD 0.276, p < 0.001) were significantly worse in the PTSD groups. Age (AUC 73.8%, p < 0.001) and EQ-5D (AUC 72.9%, p < 0.001) were independent factors that were predictive of PTSD in patients undergoing THA and TKA, respectively. When adjusting for confounding variables, PTSD was clinically and statistically significantly (p < 0.001) associated with a lower improvement in the Oxford (THA:MD 9.3; TKA:MD 10.0) and EQ-5D (THA:MD 0.375; TKA:MD 0.293) scores. CONCLUSIONS: One in 25 patients met a provisional PTSD diagnosis; they were younger and had worse preoperative and improvement in postoperative joint specific function and HRQoL. Age and EQ-5D could be used to identify patients at risk.

Age-gender differences in the relationships between physical and mental health.

Previous studies have identified that smoking, exercise and breadth of social interaction mediate the strong associations between physical and mental health. However, these studies have been restricted to older populations, have not explored differences by gender, and have not considered online social interaction. We explore how the effects of four mediators (exercise, smoking, in-person and online social interaction) of the two-way relationships between past and future physical and mental health vary across eight age and gender groups. We use data from a representative sample of the UK population consisting of 175,779 observations on 41,995 adults from Understanding Society (UKHLS) between 2009 and 2019. Within a mediation framework, we estimate the percentage of the total effects that can be explained by the proposed mediating factors. We show that exercise, smoking, in-person and online social interaction are significant mediators of the effect of mental health on future physical health. In-person social interaction is the largest of these, accounting for 2.3% of the total effect. Smoking, in-person and online interaction are significant mediators of the effect of physical health on future mental health. Again, in-person interaction is the largest of these, accounting for 3.0% of the total effect. The percentages of the total effects mediated by each factor differ substantially by age and gender. Seeking to avoid the harmful effects of poor physical health on future mental health should focus on increasing physical activity in older men, and on increasing in-person social interaction in both men and women. Seeking to avoid the harmful effects of poor mental health on future physical health should focus on increasing physical activity and in-person social interaction in older men and women, and on reducing smoking in younger men and women.

Gastrointestinal parasites in captive olive baboons in a UK safari park.

From the safety inside vehicles, Knowsley Safari offers visitors a close-up encounter with captive olive baboons. As exiting vehicles may be contaminated with baboon stool, a comprehensive coprological inspection was conducted to address public health concerns. Baboon stools were obtained from vehicles, and sleeping areas, inclusive of video analysis of baboon­vehicle interactions. A purposely selected 4-day sampling period enabled comparative inspections of 2662 vehicles, with a total of 669 baboon stools examined (371 from vehicles and 298 from sleeping areas). As informed by our pilot study, front-line diagnostic methods were: QUIK-CHEK rapid diagnostic test (RDT) (Giardia and Cryptosporidium), Kato­Katz coproscopy (Trichuris) and charcoal culture (Strongyloides). Some 13.9% of vehicles were contaminated with baboon stool. Prevalence of giardiasis was 37.4% while cryptosporidiosis was <0.01%, however, an absence of faecal cysts by quality control coproscopy, alongside lower than the expected levels of Giardia-specific DNA, judged RDT results as misleading, grossly overestimating prevalence. Prevalence of trichuriasis was 48.0% and strongyloidiasis was 13.7%, a first report of Strongyloides fuelleborni in UK. We advise regular blanket administration(s) of anthelminthics to the colony, exploring pour-on formulations, thereafter, smaller-scale indicator surveys would be adequate.

Molecular Epidemiology and Assemblage Typing of Giardia duodenalis in School-Age Children Situated along the Southern Shoreline of Lake Malawi, Malawi.

Almost all human giardiasis infections are caused by Giardia duodenalis assemblages A and B. Differentiation between human infections with these assemblages, as well as between single-assemblage (A or B) and mixed-assemblage (A and B) infections, is therefore needed to better understand the pathological impact of infection with either, or both, assemblages. We assessed the prevalence of G. duodenalis assemblages A and B using 305 fecal samples provided by school-age children situated along the southern shoreline of Lake Malawi. Concurrently, intestinal pathology data were also collected to test for association(s) between assemblage infection status and intestinal health. Prevalence of G. duodenalis infection was 39.3% by real-time polymerase chain reaction. Of all identified infections, 32% were single G. duodenalis assemblage A and 32% were single G. duodenalis assemblage B, whereas 33% were mixed-assemblage infections. Fifteen unique G. duodenalis assemblage A and 13 unique G. duodenalis assemblage B ß-giardin haplotypes were identified. There was a positive association between single infection with G. duodenalis assemblage B and both self-reporting of abdominal pain (odds ratio [OR]: 3.05, P = 0.004) and self-reporting of diarrhea (OR: 3.1, P = 0.003). No association between single infection with assemblage A and any form of intestinal pathology was found. Additionally, there was a positive association between mixed-assemblage infections and self-reporting of abdominal pain (OR: 3.1, P = 0.002). Our study highlights the importance G. duodenalis assemblage typing and reaffirms the need for improved access to water, sanitation and hygiene infrastructure in rural areas of low- and middle-income countries.

ON LIVER FLUKE (FASCIOLA HEPATICA) IN CAPTIVE VICUÑAS (VICUGNA VICUGNA) AT KNOWSLEY SAFARI, PRESCOT, UNITED KINGDOM.

Knowsley Safari (KS), Prescot, United Kingdom houses a variety of captive exotic ungulates. As part of their animal welfare plan, a prospective coprological survey was undertaken for liver fluke. In June 2021, 330 fecal samples, representative of 18 exotic ungulate species, were processed by sedimentation and filtration, with examination by coproscopy. Finding fascioliasis in all five vicuña alone, with fecal egg counts ranging from one to eight eggs per gram, anthelminthic treatment was attempted twice, with three coprological reviews. While the first anthelminthic treatment (oxyclozanide) was equivocal, the second anthelminthic treatment (triclabendazole) was proven effective upon two later follow-ups. An initial malacological survey of 16 freshwater sites in KS, first found Galba truncatula at two sites in June 2021, then upon more extensive searching subsequently within the vicuña's enclosure. It appears that F. hepatica was locally acquired, being the first report of fascioliasis within captive vicuñas in the United Kingdom. To develop a better fluke-management plan, regular coprological and malacological surveillance is justified, perhaps with molecular xenomonitoring of snails, alongside prompt administration of appropriate flukicide as required.

A systematic review of non-coding RNA genes with differential expression profiles associated with autism spectrum disorders.

AIMS: To identify differential expression of shorter non-coding RNA (ncRNA) genes associated with autism spectrum disorders (ASD). BACKGROUND: ncRNA are functional molecules that derive from non-translated DNA sequence. The HUGO Gene Nomenclature Committee (HGNC) have approved ncRNA gene classes with alignment to the reference human genome. One subset is microRNA (miRNA), which are highly conserved, short RNA molecules that regulate gene expression by direct post-transcriptional repression of messenger RNA. Several miRNA genes are implicated in the development and regulation of the nervous system. Expression of miRNA genes in ASD cohorts have been examined by multiple research groups. Other shorter classes of ncRNA have been examined less. A comprehensive systematic review examining expression of shorter ncRNA gene classes in ASD is timely to inform the direction of research. METHODS: We extracted data from studies examining ncRNA gene expression in ASD compared with non-ASD controls. We included studies on miRNA, piwi-interacting RNA (piRNA), small NF90 (ILF3) associated RNA (snaR), small nuclear RNA (snRNA), small nucleolar RNA (snoRNA), transfer RNA (tRNA), vault RNA (vtRNA) and Y RNA. The following electronic databases were searched: Cochrane Library, EMBASE, PubMed, Web of Science, PsycINFO, ERIC, AMED and CINAHL for papers published from January 2000 to May 2022. Studies were screened by two independent investigators with a third resolving discrepancies. Data was extracted from eligible papers. RESULTS: Forty-eight eligible studies were included in our systematic review with the majority examining miRNA gene expression alone. Sixty-four miRNA genes had differential expression in ASD compared to controls as reported in two or more studies, but often in opposing directions. Four miRNA genes had differential expression in the same direction in the same tissue type in at least 3 separate studies. Increased expression was reported in miR-106b-5p, miR-155-5p and miR-146a-5p in blood, post-mortem brain, and across several tissue types, respectively. Decreased expression was reported in miR-328-3p in bloods samples. Seven studies examined differential expression from other classes of ncRNA, including piRNA, snRNA, snoRNA and Y RNA. No individual ncRNA genes were reported in more than one study. Six studies reported differentially expressed snoRNA genes in ASD. A meta-analysis was not possible because of inconsistent methodologies, disparate tissue types examined, and varying forms of data presented. CONCLUSION: There is limited but promising evidence associating the expression of certain miRNA genes and ASD, although the studies are of variable methodological quality and the results are largely inconsistent. There is emerging evidence associating differential expression of snoRNA genes in ASD. It is not currently possible to say whether the reports of differential expression in ncRNA may relate to ASD aetiology, a response to shared environmental factors linked to ASD such as sleep and nutrition, other molecular functions, human diversity, or chance findings. To improve our understanding of any potential association, we recommend improved and standardised methodologies and reporting of raw data. Further high-quality research is required to shine a light on possible associations, which may yet yield important information.

Seasonality of the Meridional Overturning Circulation in the subpolar North Atlantic.

Understanding the variability of the Atlantic Meridional Overturning Circulation is essential for better predictions of our changing climate. Here we present an updated time series (August 2014 to June 2020) from the Overturning in the Subpolar North Atlantic Program. The 6-year time series allows us to observe the seasonality of the subpolar overturning and meridional heat and freshwater transports. The overturning peaks in late spring and reaches a minimum in early winter, with a peak-to-trough range of 9.0 Sv. The overturning seasonal timing can be explained by winter transformation and the export of dense water, modulated by a seasonally varying Ekman transport. Furthermore, over 55% of the total meridional freshwater transport variability can be explained by its seasonality, largely owing to overturning dynamics. Our results provide the first observational analysis of seasonality in the subpolar North Atlantic overturning and highlight its important contribution to the total overturning variability observed to date.

Biomphalaria pfeifferi (Gastropoda: Planorbidae) in Lake Malawi and Upper Shire River, Mangochi District, Malawi: Distribution, Genetic Diversity and Pre-Patent Schistosome Infections.

In November 2017, Biomphalaria pfeifferi, the key intermediate host for Schistosoma mansoni in Africa, was first reported in Lake Malawi, Mangochi District. Two subsequent malacological surveys in 2018 and 2019 confirmed its lacustrine presence, as well as its presence along the Upper Shire River. These surveys provided sufficient specimens for analyses of the genetic structure and a transmission assessment for intestinal schistosomiasis. A total of 76 collected snails were characterized by a DNA sequence analysis of a 650 bp fragment of the mitochondrial cytochrome oxidase subunit 1 (cox1); by size fractionation of six fluorescently labelled microsatellite loci (Bgµl16, Bgµl, Bpf8, rg6, U-7, and rg9);by denaturing PAGE; and by detection of pre-patent Schistosoma infection by real-time PCR with a TaqMan® probe. Five closely related cox1 haplotypes were identified, all present within a single location, with only one haplotype common across all the other locations sampled. No allelic size variation was detected with the microsatellites and all loci were monomorphic. Overall, the pre-patent prevalence of Schistosoma spp. was 31%, with infected snails found at several sampling locations. In this part of Lake Malawi, Bi. pfeifferi exhibits low genetic diversity and is clearly being exposed to the miracidia of S. mansoni, which is likely facilitating the autochthonous transmission of this parasite.

Human theca arises from ovarian stroma and is comprised of three discrete subtypes.

Theca cells serve multiple essential functions during the growth and maturation of ovarian follicles, providing structural, metabolic, and steroidogenic support. While the function of theca during folliculogenesis is well established, their cellular origins and the differentiation hierarchy that generates distinct theca sub-types, remain unknown. Here, we performed single cell multi-omics analysis of primary cell populations purified from human antral stage follicles (1-3 mm) to define the differentiation trajectory of theca/stroma cells. We then corroborated the temporal emergence and growth kinetics of defined theca/stroma subpopulations using human ovarian tissue samples and xenografts of cryopreserved/thawed ovarian cortex, respectively. We identified three lineage specific derivatives termed structural, androgenic, and perifollicular theca cells, as well as their putative lineage-negative progenitor. These findings provide a framework for understanding the differentiation process that occurs in each primordial follicle and identifies specific cellular/molecular phenotypes that may be relevant to either diagnosis or treatment of ovarian pathologies.

Rapid identification of mosquito species and age by mass spectrometric analysis.

BACKGROUND: A rapid, accurate method to identify and to age-grade mosquito populations would be a major advance in predicting the risk of pathogen transmission and evaluating the public health impact of vector control interventions. Whilst other spectrometric or transcriptomic methods show promise, current approaches rely on challenging morphological techniques or simple binary classifications that cannot identify the subset of the population old enough to be infectious. In this study, the ability of rapid evaporative ionisation mass spectrometry (REIMS) to identify the species and age of mosquitoes reared in the laboratory and derived from the wild was investigated. RESULTS: The accuracy of REIMS in identifying morphologically identical species of the Anopheles gambiae complex exceeded 97% using principal component/linear discriminant analysis (PC-LDA) and 84% based on random forest analysis. Age separation into 3 different age categories (1 day, 5-6 days, 14-15 days) was achieved with 99% (PC-LDA) and 91% (random forest) accuracy. When tested on wild mosquitoes from the UK, REIMS data could determine the species and age of the specimens with accuracies of 91 and 90% respectively. CONCLUSIONS: The accuracy of REIMS to resolve the species and age of Anopheles mosquitoes is comparable to that achieved by infrared spectroscopy approaches. The processing time and ease of use represent significant advantages over current, dissection-based methods. Importantly, the accuracy was maintained when using wild mosquitoes reared under differing environmental conditions, and when mosquitoes were stored frozen or desiccated. This high throughput approach thus has potential to conduct rapid, real-time monitoring of vector populations, providing entomological evidence of the impact of alternative interventions.

Understanding the effects of ball orientation in Rugby Union place kicking: the preferences of international kickers and the kinematics of the foot-ball impact.

Rugby Union place kicking is influential to match outcome. Previous research has analysed kicker motion prior to ball contact in detail, but ball orientation and the impact phase are typically ignored. This study aims to firstly identify the ball orientations used by international place kickers, and secondly to experimentally analyse the foot-ball interaction in trained kickers using different ball orientations. Overall, 25.5% of the international kickers used an upright ball orientation, 27.5% used a diagonal orientation and 47.1% used a horizontal orientation. However, ball orientation preference was not significant in predicting kick outcome in a binomial logistic regression model. To address the second aim, ball orientation was experimentally manipulated and lower limb and ball kinematics were captured using high-speed (4000 Hz) video. Whilst the impact location on the ball differed significantly between most ball orientation conditions, the impact location relative to the global vertical was largely consistent across all conditions. This was likely due to kickers adopting very consistent lower limb kinematics, although the shank and ankle angles at impact were affected by ball orientation conditions for some kickers. Impact durations also differed between some conditions, although this did not appear to affect the impact efficiency.