Refugia, climatic conditions and farm management factors as drivers of adaptation in Nematodirus battus populations.

Parasites are known for their ability to rapidly adapt to changing conditions. For parasitic helminths, changes in climate, along with farming and management practices associated with the intensification of livestock farming, provide novel challenges which can impact on their epidemiology and control. The sustainability of livestock production partially relies on effective control of helminth infection. Therefore, understanding changes in parasite behaviour, and what drives these, is of great importance. Nematodirus battus is an economically important helminth in the UK and temperate regions. Its infective larvae typically overwinter in eggs on pasture and hatch synchronously in spring, causing acute disease in lambs. Attempts to control disease typically rely on whole-flock benzimidazole (BZ) treatments. In recent years, the emergence of BZ-resistance, alongside the hatching of eggs without the classical over-winter 'chill stimulus', have made N. battus more difficult to control. In three previous studies, after collecting a large number of N. battus populations alongside farm management data from commercial farms, we explored the prevalence of genetic mutations associated with BZ-resistance (n = 253 farms), the ability of eggs to hatch with and without a chill stimulus (n = 90 farms) and how farm management practices varied throughout the UK (n = 187 farms). In the present study, we identify factors which may be acting as drivers, or barriers, to either the development of resistance or the variable hatching behaviour of N. battus eggs. Generalised linear mixed effect models were applied to regress experimental hatching and genotyping data on farm management and additional environmental data. Both variable hatching and resistance development appeared associated with the maintenance of parasite refugia as well as grazing management, particularly reseeding of pasture routinely grazed by young lambs each spring and the practice of set-stocked grazing. Effective quarantine measures were identified as the main protective factor for the development of BZ-resistance whereas set stocked grazing and population bottlenecks, resulting from reseeding heavily contaminated pastures, were risk factors. Spring maximum temperature and other climatic factors were associated with 'typical' hatching of eggs following a chill stimulus whilst several management factors were linked with hatching without prior chilling. For example, practices which reduce parasite numbers on pasture (e.g. re-seeding) or restrict availability of hosts (e.g. resting fields), were found to increase the odds of non-chill hatching. Retention of the timing of lambing and infection level of the host within the fitted model indicated that requirement for a chill stimulus prior to hatching may be plastic, perhaps subject to change throughout the grazing season, in response to immune development or parasite density-dependence within the host. Further investigation of the influence of the factors retained within the fitted models, particularly the theme of parasite refugia which was highlighted in relation to both the presence of BZ-resistance alleles and alternative hatching, is required to establish robust, sustainable parasite control and farm management strategies.

Artificial Intelligence in Ultrasound Diagnoses of Ovarian Cancer: A Systematic Review and Meta-Analysis.

Ovarian cancer is the sixth most common malignancy, with a 35% survival rate across all stages at 10 years. Ultrasound is widely used for ovarian tumour diagnosis, and accurate pre-operative diagnosis is essential for appropriate patient management. Artificial intelligence is an emerging field within gynaecology and has been shown to aid in the ultrasound diagnosis of ovarian cancers. For this study, Embase and MEDLINE databases were searched, and all original clinical studies that used artificial intelligence in ultrasound examinations for the diagnosis of ovarian malignancies were screened. Studies using histopathological findings as the standard were included. The diagnostic performance of each study was analysed, and all the diagnostic performances were pooled and assessed. The initial search identified 3726 papers, of which 63 were suitable for abstract screening. Fourteen studies that used artificial intelligence in ultrasound diagnoses of ovarian malignancies and had histopathological findings as a standard were included in the final analysis, each of which had different sample sizes and used different methods; these studies examined a combined total of 15,358 ultrasound images. The overall sensitivity was 81% (95% CI, 0.80-0.82), and specificity was 92% (95% CI, 0.92-0.93), indicating that artificial intelligence demonstrates good performance in ultrasound diagnoses of ovarian cancer. Further prospective work is required to further validate AI for its use in clinical practice.

Laboratory diagnosis of gastrointestinal nematodes in first-grazing-season cattle.

This focus article has been prepared by Natalie Jewell of the APHA Cattle Expert Group with the assistance of Vanessa Swinson (veterinary lead of the APHA Cattle Expert Group), Claire Hayman, Lucy Martindale and Anna Brzozowska (Surveillance Intelligence Unit), and Siân Mitchell (formerly the APHA's parasitology discipline champion).

Is the Ollerenshaw fasciolosis forecasting model fit for the 21st century?

BACKGROUND: The Ollerenshaw forecasting model is based on rainfall and evapotranspiration and has been in use to predict losses from fasciolosis since 1959. We evaluated the performance of the model against observed data. METHODS: Weather data were used to calculate, map and plot fasciolosis risk values for each year from 1950 to 2019. We then compared the model's predictions with recorded acute fasciolosis losses in sheep from 2010 to 2019 and calculated the sensitivity and specificity of the model. RESULTS: The forecast risk has varied over time but has not markedly increased over the past 70 years. The model correctly forecasted the highest and lowest incidence years at both the regional and national (Great Britain) levels. However, the sensitivity of the model for predicting fasciolosis losses was poor. Modification to include the full May and October rainfall and evapotranspiration values made only a small improvement. LIMITATIONS: Reported acute fasciolosis losses are subject to bias and error due to unreported cases and variations in region size and livestock numbers. CONCLUSION: The Ollerenshaw forecasting model, in either its original or modified forms, is insufficiently sensitive to be relied upon as a standalone early warning system for farmers.

Practical guide to the diagnostics of ruminant gastrointestinal nematodes, liver fluke and lungworm infection: interpretation and usability of results.

The diagnostics of ruminant parasites remains one of the cornerstones for parasite control best practices. Field veterinarians have several techniques at their disposal (fecal egg count, coproculture, FAMACHA®, plasma pepsinogen, ELISA-Ostertagia, ELISA-Fasciola, Baermann and ELISA-Lungworm) for the identification and/or quantification of gastrointestinal nematodes, lungworms and liver fluke infecting small ruminants and cattle. Each of these diagnostic tools has its own strengths and weaknesses and is more appropriate for a specific production operation and/or age of the animal (young and adults). This review focuses on the usability and interpretation of the results of these diagnostic tools. The most advanced technical information on sampling, storage, advantages and limitations of each tool for different types of production operations and animal categories is provided.

Perinatal outcomes in women with sickle cell disease: a matched cohort study from London, UK.

There are limited data on contemporary outcomes for women with sickle cell disease (SCD) in pregnancy. We conducted a single-site matched cohort study, comparing 131 pregnancies to women with SCD between 2007 and 2017 to a comparison group of 1310 pregnancies unaffected by SCD. Restricting our analysis to singleton pregnancies that reached 24 weeks of gestation, we used conditional Poisson regression to estimate adjusted risk ratios (aRRs) for perinatal outcomes. Infants born to mothers with SCD were more likely to be small for gestational age [aRR 1·69, 95% confidence interval (CI) 1·13-2·48], preterm (aRR 2·62, 95% CI 1·82-3·78) and require Neonatal Unit (NNU) admission (aRR 3·59, 95% CI 2·18-5·90). Pregnant women with SCD were at higher risk of pre-eclampsia/eclampsia (aRR 3·53, 95% CI 2·00-6·24), more likely to receive induction of labour (aRR 2·50, 95% CI 1·82-1·76) and caesarean birth (aRR 1·44, 95% CI 1·18-1·76). In analysis stratified by genotype, the risk of adverse outcomes was highest in haemoglobin SS (HbSS) pregnancies (n = 80). There was no strong evidence that haemoglobin SC (HbSC) pregnancies (n = 46) were at higher risk of preterm birth, caesarean delivery, or NNU admission. Pre-eclampsia/eclampsia was more frequently observed in HbSC pregnancies. Despite improvements in the care of pregnant women with SCD, the increased risk of adverse perinatal outcomes remains.

Exploration of factors associated with spatial-temporal veterinary surveillance diagnoses of rumen fluke (Calicophoron daubneyi) infections in ruminants using zero-inflated mixed modelling.

Rumen fluke (Calicophoron daubneyi) has emerged as a prominent parasite of ruminants in Europe over the past decades. Epidemiological questions remain regarding this observed increase in prevalence as well as the prospect for future paramphistomosis risk. This study aimed to identify factors associated with the temporal−spatial prevalence of rumen fluke as measured by veterinary surveillance in a temperate region using zero-inflated negative binomial mixed modelling. Modelling revealed that summer rainfall, raindays and sunshine hours and mean winter temperature as significant positively associated climate variables for rumen fluke prevalence over space and time (P < 0.05). Rumen fluke prevalence was also higher in counties with higher cattle/sheep densities and was positively associated with rumen fluke case rates in the previous years (P < 0.05). Equivalent models for fasciolosis prevalence revealed no significant association with winter temperature and sunshine hours, (P > 0.05). These results confirm a strong association between rainfall and the prevalence of both fluke species in a temperate environment, likely due to the role of Galba truncatula as their intermediate snail host. It also highlights the potential added importance of winter temperature and sunshine hours in rumen fluke epidemiology when compared to liver fluke.

Descriptive analysis of nematode management practices and Nematodirus battus control strategies on UK sheep farms.

BACKGROUND: Farm management practices have a major impact on nematode population dynamics. The presented study aimed to understand current nematode management practices on UK sheep farms; with a particular focus on Nematodirus battus because of the changing epidemiology, and emerging anthelmintic resistance observed in this species. METHODS: A 42 question online survey covering grazing management, farm demographics and parasite control strategies was developed and distributed to the farming community in 2016. Analysis of the 187 completed questionnaires explored regional variations in practices. RESULTS: Uptake of recommendations was variable, particularly quarantine practices and monitoring tools. Results also highlighted variation in the epidemiology of N. battus; respondents in the north (Scotland, north-west and north-east England) typically reported N. battus in spring with a perception of more severe clinical symptoms than those from the south (Midlands, Wales, south-east and south-west England; p = 0.03). Farms in the south observed greater changes in the timing of disease (p = 0.006) with N. battus being reported throughout the year on some holdings and more frequent use of faecal egg count monitoring (p = 0.006). CONCLUSIONS: Control of N. battus infection is challenging and 'one-size-fits-all' advice is not applicable; however, the information gathered will enable the development of effective, adaptable control strategies.

Rumen Fluke in Great Britain.

Calicophoron daubneyi is the primary rumen fluke (RF) found in Europe in ruminants and infection is more common in cattle than in sheep. The incidence of RF has appeared to increase greatly throughout Europe in the last 10-15 years, with outbreaks of clinical paramphistomosis confirmed in ruminants in many countries, including Great Britain and Ireland. Clinical disease, due to immature stages developing in the small intestine, appears infrequently but can occur, usually in the autumn or winter within weeks of beginning to graze wet pasture. Although disease due to adult RF has not been proven, subclinical production losses have been attributed to adult RF infection by some researchers. As the intermediate host for RF and the liver fluke (Fasciola hepatica) is the mud snail (Galba truncatula), similar habitats and environmental conditions favour both parasites. There may, however, be differences in parasite development and interactions within both the final and intermediate hosts. No anthelminthic product is licensed for treatment of ruminants for RF in the UK. However, oxyclozanide, licensed for the treatment of adult F. hepatica infection, has been shown to have activity, but it may be more effective against the adult than the immature stages. The future prevalence of RF due to climate change and limited treatment options is unpredictable. Infection and clinical disease could become more common and RF is worthy of further research.

Molecular and phenotypic characterisation of fenbendazole resistance in a field-derived isolate of Ostertagia ostertagi.

The prevalence of anthelmintic resistance in the bovine nematode Cooperia oncophora has been well documented globally but lack of efficacy against the more pathogenic nematode species Ostertagia ostertagi is less common. The sensitivity of an O. ostertagi isolate to the benzimidazole class of anthelmintic was investigated using classical parasitological techniques following apparent clinical failure of controlled release fenbendazole capsule administration in first season grazers at pasture. A controlled efficacy test (CET) was conducted in conjunction with sequencing of the ß-tubulin isotype 1 gene of larvae pre- and post-fenbendazole administration. Twelve helminth-naïve calves were infected experimentally with 20,000 third stage larvae; six received oral fenbendazole (7.5 mg/kg bodyweight) 28 days post infection. Total abomasal nematode burdens were compared between treatment and control groups to determine efficacy. Fenbendazole resistance in O. ostertagi was confirmed with a total treatment failure in reducing worm burden: efficacy of 0%. Sequence analysis of the ß-tubulin isotype-1 gene from forty-five infective larvae from both control and treated groups was performed. The three commonest single nucleotide polymorphisms (SNPs) associated with benzimidazole resistance, namely F167Y, E198A and F200Y, were examined. The predominant resistance-associated SNPs were F200Y (78 % control and 79 % treated groups) and F167Y (remaining genotypes) and emphasises the importance of these SNPs in clinical disease in this isolate. The development of diagnostic molecular tools based on a characterised field-derived isolate of benzimidazole-resistant Ostertagia will enable future prevalence surveys to be undertaken to assess the possible risk posed by resistance in this economically important species.

Variation in hatching responses of Nematodirus battus eggs to temperature experiences.

BACKGROUND: Nematodirus battus, unlike most other gastrointestinal nematodes, undergoes maturation to an infective larva within the egg. Historically, eggs were considered to require a period of chilling over winter followed by a period of temperature above 10 °C for synchronous hatching to occur (generally in spring). Anecdotal reports of Nematodirus infection out-with spring in veterinary journals and the farming press suggest that the concentrated pasture abundance of N. battus infective larvae may be changing. In order for control practices to be adapted, and unexpected disease outbreaks to be avoided, it is important to quantify how parasite epidemiology is changing and research the drivers behind it. METHOD: The present study investigated the in vitro hatching response to temperature experiences (with and without a period of chilling) for egg samples of 90 N. battus populations obtained from 73 commercial sheep farms. Six aliquots of larvated eggs were prepared per population, three aliquots were placed at 4 °C for 6  weeks to provide a chill stimulus then incubated at the optimal hatching temperature for the species. The remaining three aliquots of eggs were incubated at the hatching temperature without a prior chill stimulus and the number of hatched larvae was compared between treatments. RESULTS: Median hatch rate across all populations with chilling was 45% (95% CI: 42-48%) and without chilling was 4% (95% CI: 2-6%). Inter-population variation in hatching ranged from 0 to 87% of eggs able to hatch in the absence of a chill stimulus, mean non-chill hatching was 13 ± 2% of eggs (mean ± SE). Non-chill hatching rates were greater than chilled hatching rates in seven of the 90 populations tested. CONCLUSIONS: Clearly, the variation in hatching responses to temperature experience is very large and therefore the seasonality of the parasite may vary not only between regions but also at farm level. In contrast to what previous work has suggested, there was a geographical trend towards higher non-chill hatching in the Northern parts of the UK.

Large scale screening for benzimidazole resistance mutations in Nematodirus battus, using both pyrosequence genotyping and deep amplicon sequencing, indicates the early emergence of resistance on UK sheep farms.

Benzimidazoles (BZ) have been the anthelmintic of choice for controlling Nematodirus battus infections since their release in the 1950s. Despite heavy reliance on this single anthelmintic drug class, resistance was not identified in this nematode until 2010 (Mitchell et al., 2011). The study aimed to explore the prevalence of BZ-resistance mutations in N. battus from UK sheep flocks using deep amplicon sequencing and pyrosequencing platforms. Based on evidence from other gastrointestinal nematodes, resistance in N. battus is likely to be conferred by single nucleotide polymorphisms (SNP) within the ß-tubulin isotype 1 locus at codons 167, 198 and 200. Pyrosequencing and deep amplicon sequencing assays were designed to identify the F167Y (TTC to TAC), E198A (GAA to GCA) and F200Y (TTC to TAC) SNPs. Nematodirus battus populations from 253 independent farms were analysed by pyrosequencing; 174 farm populations were included in deep amplicon sequencing and 170 were analysed using both technologies. F200Y was the most prevalent SNP identified throughout the UK, in 12-27% of the populations tested depending on assay, at a low overall individual frequency of 2.2 ±â€¯0.6% (mean ±â€¯SEM, based on pyrosequencing results). Four out of the five populations with high frequencies (>20%) of the F200Y mutation were located in NW England. The F167Y SNP was identified, for the first time in this species, in four of the populations tested at a low frequency (1.2% ±â€¯0.01), indicating the early emergence of the mutation. E198A or E198L were not identified in any of the isolates. Results obtained were comparable between both techniques for F200Y (Lins' CCC, rc = 0.96) with discrepancies being limited to populations with low frequencies. The recent emergence of resistance in this species will provide a unique opportunity to study the early stages of anthelmintic resistance within a natural setting and track its progress in the future.

First detection of ivermectin resistance in oesophagostomum dentatum in pigs.

In 2016 suspected reduced ivermectin (IVM) efficacy in Oesophagostomum species in pigs was reported in England. Following this initial report, APHA raised awareness amongst private pig veterinary practitioners of the need to monitor the efficacy of the worm control on pig units. In 2017 another veterinary practitioner highlighted a potential in-field lack of IVM efficacy in treating Oesophagostomum species in sows on another breeder-finisher unit. In this trial, the efficacy of IVM against Oesophagostomum species worms has been investigated to determine whether suspected reduced efficacy (52% reduction in mean faecal egg count 14 days post ivermectin administration) on a mixed indoor and outdoor breeder-finisher pig farm in England reflected true IVM resistance under controlled experimental conditions. On days 0 and 40 of the trial, twenty helminth-naive pigs were artificially infected per os with 5000 Oesophagostomum L3 obtained from the farm under investigation. The pigs were allocated to treatment or control groups (n = 10 per group). Treatment group pigs received IVM (0.3 mg kg body weight) by sub-cutaneous injection as per manufacturer's instructions on day 44. Control group animals were left untreated. Faecal worm egg counts were monitored throughout the trial from day 15 post infection to determine time to patency. On day 50 all pigs were euthanased to assess the worm burdens. Resistance to IVM was confirmed in Oesophagostomum dentatum based on the results of a faecal egg count reduction test (FECRT) and a controlled efficacy test (CET). Efficacy based on mean reduction in faecal egg count of IVM-treated pigs compared to untreated control pigs was 86%. Mean reduction in IVM-treated pig worm burdens was 5% against an adult worm population and 94% against an L3/L4 population. The apparent discrepancy between FECRT and CET efficacy results appears to be due to egg development and/or oviposition suppression in IVM-treated female worms. The detection of IVM resistance in Oesophagostomum species worms for the first time in UK pigs is particularly important considering the global situation where resistance to pyrantel, levamisole and benzimidazole anthelmintics in Oesophagostomum species in pigs have already been reported. The results also provide an opportunity to discuss the wider issue of anthelmintic usage and efficacy on pig farms and highlight the need for wider surveillance for the occurrence of anthelmintic resistance in pigs.

100 Questions in Livestock Helminthology Research.

An elicitation exercise was conducted to collect and identify pressing questions concerning the study of helminths in livestock, to help guide research priorities. Questions were invited from the research community in an inclusive way. Of 385 questions submitted, 100 were chosen by online vote, with priority given to open questions in important areas that are specific enough to permit investigation within a focused project or programme of research. The final list of questions was divided into ten themes. We present the questions and set them briefly in the context of the current state of knowledge. Although subjective, the results provide a snapshot of current concerns and perceived priorities in the field of livestock helminthology, and we hope that they will stimulate ongoing or new research efforts.

First evidence of resistance to macrocyclic lactones in Psoroptes ovis sheep scab mites in the UK.

Ovine psoroptic mange (sheep scab) is an infection of substantial economic and animal welfare concern in the UK. Its prevalence has increased rapidly over the last 20 years and management is dependent on a small number of acaricidal compounds, many of which are also used to control a range of other endoparasites and ectoparasites. Here, the effects of the macrocyclic lactone (ML) moxidectin was considered using in vitro assays against mites from four farm populations where persistent treatment failure had been reported: two in West Wales, one from the England/Wales border and one in Herefordshire. The data demonstrate resistance in mites from all four farms. This is the first quantitative evidence of ML resistance in Psoroptes mites in the UK. Given the similarities in their mode of action it is highly likely that cross-resistance across the range of this class of compound will be found. The development of resistance to moxidectin is of considerable concern given the already high prevalence of scab infection in some regions; major difficulties in scab management should be anticipated if ML resistance becomes widely established in the UK.

Aspergillus niger infection in an immunosuppressed patient confined solely to the brain.

A 68-year-old woman with a background of hypertension, stroke and rheumatoid arthritis presented to her local hospital after a 4-week history of gradual deterioration and increasing confusion with new onset right-sided weakness. Her initial CT scan revealed a rim enhancing mass lesion with surrounding oedema in the left parietal lobe for which she underwent CT stealth-guided biopsy. Microbiology culture of the 2 biopsy samples yielded Aspergillus niger and she was started on the antifungal agent voriconazole. MRI 2 weeks after the procedure also demonstrated radiological findings consistent with intracranial aspergillosis. She later developed leucopenia with neutrophils of 1.5×109/L and her methotrexate and voriconazole were stopped. Voriconazole was changed to oral posaconazole. She did not undergo surgical resection and has continued to improve clinically on posaconazole, with recovery in her white cell count.