Conference Agenda

Many parasites have elaborate life cycles with multiple stages and strict host requirements. The biological questions posed by these life cycles are fascinating, but their complexity can be overwhelming and showing every detail sends audiences reaching for their phones. As a result, life cycles are frequently overlooked, even though they are fundamental to questions of why parasites live the way they do, how they adapt to changing environments, and whether we can exploit life cycle traits to eradicate disease.

We aim to answer some of these questions by studying the mosquito stages of the malaria life cycle. To survive in the insect host, malaria parasites must adapt to a radically different physical and immunological environment. They do this by rapidly and irreversibly progressing through distinct morphological forms and significantly altering their metabolic processes. Faced with these extreme changes, the parasite population shrinks dramatically, and the surviving parasites grow very slowly. Somewhat surprisingly, parasites complete their obligate sexual reproduction during this period of stress and parasite death. Recent advancers in our understanding of the biological processes and genetic impacts of mosquito-stage development highlight significant vulnerabilities that can be exploited to disrupt disease transmission. We’ve identified anti-malarial compounds that can be delivered across the mosquito life-stages to directly kill parasites with minimal selection for resistance, and drug-resistant parasites with little or no ability to survive under the metabolic demands of the mosquito stages. We’re also exploring new genetic tools than can be exploit the obligate sexual stages to spread through and modify entire parasite populations.

Surprisingly, we discovered significant gaps in our knowledge of the well-studied malaria parasite life cycle. Being able to fill these gaps is an important benefit of developing new anti-malarial strategies and highlights the practical importance of thoroughly understanding parasite life cycles. While the details of these new anti-parasite strategies are malaria specific, we believe that many other parasite life cycles share these vulnerabilities and can be targeted using similar approaches tailored to specific parasite biology.

Mammalian meat allergy (MMA) is an IgE-mediated allergic condition triggered by sensitisation to galactose-α-1,3-galactose (α-Gal) following tick bite. First described in Australia in 2007, MMA is now recognised globally, yet its national burden in Australia has never been systematically characterised. We addressed this gap by analysing 11 years of national α-Gal specific IgE testing data (2014–2024), encompassing over 16,000 tests from 14,000 individuals. Our findings reveal MMA as a substantial and rapidly growing health concern, with case detection increasing 22% annually since 2020. The geographic distribution of cases closely mirrors the range of Ixodes holocyclus, with extreme spatial clustering within this endemic zone: just nine discrete regions account for over half od the national MMA disease burden. This tight coupling between vector ecology and disease distribution underscores the central role of I. holocyclus in driving MMA in Australia, with only minor contributions from other tick species. Among serially tested patients, α-Gal specific IgE levels declined over time in the vast majority of individuals, supporting the utility of repeat testing for clinical monitoring. This study provides the first comprehensive national epidemiological assessment of MMA in Australia and establishes baseline metrics for ongoing surveillance of this emerging tick-borne allergic condition.

Late-arresting sporozoite vaccines against Plasmodium falciparum achieve high efficacy but pose manufacturing and intravenous delivery challenges. We describe an alternative chemo-attenuation strategy that exploits first-in-class antimalarials targeting parasite aspartyl proteases plasmepsin IX and X. A single low-dose infection with virulent Plasmodium berghei sporozoites, delivered intravenously or by mosquito bite, followed by cure with a dual plasmepsin IX/X inhibitor prevented blood-stage infection by generating chemo-attenuated liver merozoites (CALM) that are incapable of erythrocyte invasion. CALM vaccination conferred sterile protection in mice for up to 2 years. Protective immunity involved both antibodies and CD8+ T cells recognising a spectrum of Plasmodium antigens including CSP, SERA1, RPL6, GAP50, RNT, PHIST, S20 and RBP. Studies in humanised mice confirmed plasmepsin IX/X inhibition similarly prevents Plasmodium falciparum liver-to-blood transition, demonstrating conservation of the drug’s late liver-stage mechanism across species. By enabling controlled liver-stage arrest without complex genetic attenuation or irradiation, plasmepsin IX/X-targeting drugs provide a practical, broadly applicable pathway to whole-parasite chemovaccination. This approach may convert seasonal mosquito exposure into progressive immune education if long-acting injectable (LAI) formulations prove feasible. These findings lay the foundation for clinical evaluation of CALM vaccination including via mosquito-bite immunisation. Aspirationally, LAI CALM could transform malaria prevention programs at scale.

The Australian sheep blowfly, Lucilia cuprina dorsalis, is a significant ectoparasite of sheep responsible for flystrike, leading to substantial production losses, animal injury, and mortality. Current flystrike control strategies rely heavily on surgical mulesing and insecticides; however, these approaches present ongoing welfare, sustainability, and efficacy challenges. Despite the economic and biological importance of this species, genomic resources for L. c. dorsalis remain limited, constraining our understanding of its biology and population dynamics. To address this gap, we employed an integrated genomics approach combining Oxford Nanopore Technologies (ONT) long-read sequencing, Illumina short-read sequencing, and Omni-C proximity ligation to generate a chromosomal-level genome assembly. Additionally, comprehensive long- and short-read RNA sequencing was used to construct a high-resolution de novo transcriptome. Together, these datasets provide a robust foundation for genome annotation and functional characterization. This multi-omics framework provides new insights into the genetic architecture, evolutionary history, and key biological processes of L. c. dorsalis. Importantly, the chromosomal-level assembly enables population genomics analyses across Australian blowfly populations, facilitating investigation of genetic diversity, structure, and dispersal. These resources establish a critical platform for advancing genomic research and supporting the development of improved and sustainable flystrike management strategies, bridging fundamental genomics with applied sheep health outcomes.

Strongyloidiasis is a much-neglected disease caused by the soil-transmitted helminth Strongyloides stercoralis, afflicting millions globally- predominantly low-income tropical regions as well as the remote Aboriginal communities of northern Australia. Current diagnostic tests for Strongyloidiasis are neither sufficiently sensitive nor field-friendly for use in low-endemic and resource-poor settings, and reliable point-of-care (POC) diagnostic tools are urgently needed for disease mapping and monitoring of control efforts.

CRISPR technologies have enabled the development of a powerful new class of rapid, ultra-sensitive, cost-effective POC diagnostics for viruses/cancers. For the first time, we developed CRISPR-based assays for the detection of Strongyloidiasis, validated using human and animal stool samples collected from S. stercoralis endemic regions. The assays were demonstrated to be highly specific, with no cross-reactivity observed with an array of bacteria/fungi/parasite species. They also showed a sensitivity comparable to qPCR (the emerging gold standard), but are more field-friendly, with significantly reduced need for specialised equipment and expertise, requiring only a simple portable heat-block and UV detection. We also developed duplex CRISPR-based assays for simultaneous detection of other major helminth infections including Schistosoma mansoni and hookworm (Necator americanus), to better investigate helminth co-infections. This CRISPR-based platform offers a promising, field-ready next-generation approach to Strongyloidiasis diagnostics.

Malaria causes significant morbidity and mortality, with 282 million cases and 610,000 deaths in 2024. The past decade has seen progress towards malaria eradication, however recent trends indicate that improvements have plateaued, partly due to parasite drug-resistance and treatment failure. To combat parasite drug resistance, new drugs with different modes of action to current antimalarials are needed. We have identified novel antiplasmodial compounds from synthetic and natural product libraries and using machine learning/AI tools. This includes the indoloquinolizidine alkaloid natural product alstonine (PfIC₅₀ of 0.18 µM, Selectivity Index (SI) >1,000), novel 1,3,4-oxadiazoles (e.g., 3 with PfIC₅₀ 0.16 µM, SI 162) and primary hydroxamates (e.g., ACY-738 with PfIC₅₀ 0.08 µM, SI 314). In this presentation, an overview will be given on strategies employed to identify these compounds and to understand their antiplasmodial action, including phenotypic analyses and using CRISPR/Cas9 mediated approaches to investigate putative targets. Elucidating the mechanism of action of antiplasmodial compounds can help identify novel druggable targets, inform downstream drug development and aid in improving our understanding Plasmodium biology.

Canine hookworms, predominantly Ancylostoma caninum, remain the most prevalent intestinal nematode of dogs in Australia, with infection dynamics strongly influenced by host age, management practices, and geographic location. Current anthelmintic control relies on three major drug classes - benzimidazoles (febantel, fenbendazole), macrocyclic lactones (moxidectin, milbemycin oxime), and tetrahydropyrimidines (pyrantel). However, an increasing number of clinical cases are characterised by persistent infections refractory to standard treatment regimens.

Recent investigations from our group provide compelling evidence that many of these cases are associated with reduced efficacy, and in some instances resistance, to fenbendazole and pyrantel. These findings raise concerns regarding the emergence of multiple anthelmintic drug resistance (MADR) in Australian A. caninum populations. Epidemiological trends indicate a disproportionate representation of affected dogs originating from breeding and training facilities, where intensive deworming and high environmental contamination likely create strong selection pressure for resistant parasite populations.

The growing movement of dogs from such facilities into the general community presents a significant risk for the dissemination of resistant hookworm strains. This concern is further amplified by recent reports of MADR A. caninum in the United States, underscoring the global relevance of this emerging threat. Adoption of evidence-based, risk-driven parasite control strategies is therefore critical to minimise unnecessary anthelmintic use and slow resistance selection.

This presentation will discuss current advances in the diagnosis of anthelmintic resistance in canine hookworms, encompassing in vitro assays, in vivo efficacy studies, and molecular approaches targeting resistance-associated markers. In addition, I will highlight the urgent need for coordinated national surveillance and reporting frameworks to detect and respond to treatment failure. Finally, the development of integrated, multidisciplinary guidelines engaging veterinarians, parasitologists, epidemiologists, and animal welfare organisations will be proposed as a strategic priority to safeguard the long-term efficacy of anthelmintics and ensure sustainable parasite control in dogs.

Helminth infections, particularly schistosomiasis and soil-transmitted helminthiases, impose a severe global health burden, infecting more than a quarter of the world population, with a disproportionate effect on those in extreme poverty. Current diagnostic tests for worm infections are neither sufficiently sensitive nor field-friendly for use in resource-limited or low-endemic settings, leading to underestimation of true infection rates. Ultrasensitive, field-friendly, low-cost point-of-care diagnostics are urgently needed to better control these diseases.
Our group has established a highly sensitive CRISPR-Cas12/13-based platform for femtogram-level nucleic acid detection of multiple helminth infections. We also developed portable, lyophilized one-pot CRISPR detection to enable field testing, reducing reliance on cold-chain transportation and making it easier to perform. To further address the DNA sample preparation bottleneck in field applications, we developed a rapid extraction method that yields high-quality DNA for use with our CRISPR-based schistosomiasis diagnostic tool. Our method is faster and supports scalable, cost-effective field screening, compared with conventional DNA extraction.
To demonstrate the clinical utility of this integrated system, we are currently validating the assay for helminth detection using a large cohort of human samples from Uganda. This scalable, field-deployable approach holds significant potential to advance global NTD control and surveillance efforts.

Giemsa-stained blood smear microscopy is the gold standard for detecting malaria parasites, but it is time-consuming and limited for storage and reference. To address this, we developed PlasmoCount, a deep learning tool for accurate, automated counting of intracellular parasites and digital archiving support. Principally, we have achieved a substantial reduction in PlasmoCount’s processing time allowing for evaluation of a single image in under 3 seconds (reduced from 40). In addition, we have updated the tool so that it can now detect blood-stage infections from multiple species of human-infective and experimental rodent-infective Plasmodium parasites. Combined with a suite of other updates, including advanced cell differentiation and use at different magnifications, these augmentations broaden the distribution of input data our model can accommodate and radically advance its speed whilst maintaining its high classification accuracy (99.8%). Finally, we provide an offline, on-device version of the standardised framework designed for smartphones, including iOS and Android operating systems. By making use of imported images or image capture via a smartphone camera, PlasmoCount 2.0 markedly improves malaria parasite smear-based detection and provides a reproducible means to assess parasite infections either in routine laboratory work or as a future aid in clinical or field diagnosis.

Countries close to malaria elimination are reporting significant resurgence following periods of declining prevalence. Reduced transmission decreases opportunities for genetic recombination and generates geographically isolated hotspots of infection, resulting in lower diversity and increased population structure. With reduced exposure, naturally acquired immunity wanes, potentially rendering human populations more susceptible to outbreaks. But paradoxically, many low-transmission countries also record high prevalence of asymptomatic infections. We hypothesised that these asymptomatic cases are associated with immunologically familiar, locally circulating strains whereas clinically infectious parasites are potentially imported. We analysed Plasmodium falciparum samples from a period of low transmission (2012) prior to resurgence (2016) in East Sepik, Papua New Guinea (PNG), and from a low-transmission setting with ongoing occupational exposure in Mondulkiri, Cambodia, by sequencing a validated genome-wide single nucleotide polymorphism (SNP) barcode and immune evasion antigen marker (varcode). Parasite lineages underlying clinical infections in PNG were clonal and distinct from circulating asymptomatic isolates, suggesting potential importation and outbreak caused by immunologically unfamiliar parasites. Infections in Cambodia however indicate a more complex dynamic between parasite strain and host factors. These results emphasise how surveillance reliant on just clinical infections inadequately reflects control success and must account for asymptomatic malaria for sustainable reduction and elimination.

Transmission is a basic aspect of intestinal parasite’s biology that is poorly understood for Dientamoeba fragilis. Early historical reports reflecting the absence of a cyst are often cited as a central argument in debates supporting the lack of a D. fragilis cyst. Despite D. fragilis cysts being described since Dobell’s original description, their existence is not universally accepted. Here, Dobell’s, Wenyon’s, and Hoare’s collection of historical faecal smears stored at the Natural History Museum (London), dating back to the 1890s and the early 1900s, were examined for forms consistent with modern descriptions of D. fragilis cysts, and an example was found in one slide. Such rare forms were also detected during examination of stained faecal smears archived in the parasite reference laboratory collection at the United States Centers for Disease Control and Prevention (CDC). Considering published literature on the subject of D. fragilis cysts and the broader picture of cyst formation across diverse members of Parabasalia, we recommended that future investigations on D. fragilis transmission consider mounting evidence for the role of a true cyst despite its rarity in human faecal specimens. The factors leading to cyst formation and further characteristics of this life cycle stage require further study.

Babesiosis, caused by intraerythrocytic parasites of the genus Babesia, is an emerging zoonotic threat with a global distribution. Babesiosis ranges from asymptomatic to fulminant disease occurring predominantly in immunocompromised hosts, with fatality rates up-to 20%. Diagnosis traditionally relies on microscopic examination of blood smears; however, up-to 20% of infections are sub‑microscopic, risking transmission via blood transfusion, organ transplantation, and the ixodid tick vector. While molecular and immunodiagnostic methods exhibit improved sensitivity, they are costly, time‑consuming, technically complex, and invasive. Near‑infrared spectroscopy (NIRS) utilises near-infrared electromagnetic energy (350–2500 nm) to generate spectral signatures reflective of specific chemical changes in a biological sample. When coupled with machine learning algorithms, diagnostic features can be extracted, allowing sample classification. NIRS has been successfully applied as a non-invasive malaria diagnostic in mice and humans; however analogous studies have not been performed with Babesia spp. This study evaluated the ability of NIRS to non-invasively detect B. microti in mice. The sensitivity and specificity of non-invasive detection was compared to invasive detection (blood spots). By situating NIRS alongside established molecular detection methods, our goal is to lay the groundwork for a rapid, reagent‑free diagnostic that complements existing assays while enabling non‑invasive babesiosis surveillance and enhanced donor screening.

Platelet Factor 4 Derived Internalisation Peptide (PDIP), based on the antimicrobial peptide-like domain of human PF4, exhibits activity against Plasmodium. PDIP rapidly kills parasites by penetrating Plasmodium-infected erythrocytes and destroying the digestive vacuole. Why PDIP penetrates only infected but not healthy erythrocytes and destroys only the digestive vacuole is unclear. We hypothesised that differences in lipid composition between infected erythrocyte and parasite membranes alter membrane polarity and fluidity, thereby facilitating PDIP’s differential interactions.

To test this, we used the membrane dyes Nile Red and Laurdan, which report polarity and fluidity through emission‑wavelength shifts quantified as ratiometric indices. These ratios were measured for uninfected and Plasmodium-infected erythrocyte membranes, and the intracellular parasite, with and without PDIP treatment. We found comparable fluidity between infected and uninfected erythrocyte membranes. Polarity differed significantly, ranking from most to least: intracellular parasite > uninfected > and infected erythrocyte membranes. These polarity differences may contribute to PDIP’s ability to selectively enter infected cells and suggested that parasites contain more polar lipids that may facilitate PDIP’s activity. PDIP treatment significantly increased polarity and reduced fluidity for all membrane types. This indicates a non-specific interaction of the peptide independent of translocation that alters membrane properties non-destructively.

Widespread resistance to all current antimalarials threatens the control and eradication of malaria. Second-generation bis-triazines display low nanomolar potency and fast-killing asexual P. falciparum. However, the novel mechanism of action (MOA) remains unknown.

In vitro combination drug-pulse assays using various inhibitors were performed to identify potential modulators of bis-triazine activity. We also included two antimalarial candidates currently under development with the Medicines for Malaria Venture (MMV) with some structural similarity to the bis-triazines. E64d, a cysteine protease inhibitor, bafilomycin A1, a V-ATPase inhibitor and chloroquine all caused antagonism of trophozoite-stage activity across the bis-triazine analogues and MMV candidates (between 2 and 10-fold increases in IC₅₀). All three activity modulators are known to localise to the digestive vacuole and indicates potential involvement of the haemoglobin digestion pathway in the MOA of these series.

3-hour ring-stage survival assays with an artemisinin-resistant clinical isolate and a Pf3D7 line genetically modified to induce knockdown of essential falcipain-3 resulted in decreased activity (up to 20-fold increases in IC₅₀) for both MMV candidates and one bis-triazine analogue. The current lead bis-triazine analogue, however, observed no change or slight hypersensitisation. Uninterrupted haemoglobin digestion appears to be vital for these compounds to maintain their fast-killing activity.

Malaria remains one of the most devastating infectious diseases globally. Resistance to frontline antimalarials continues to compromise control efforts, highlighting the urgent need for novel therapeutic strategies. We previously identified the strobilurin compound MMV1794211 as a highly potent antiplasmodial agent with low-picomolar activity against blood-stage Plasmodium falciparum. Biochemical and enzymatic analyses demonstrated that MMV1794211 targets Complex III of the parasite mitochondrial electron transport chain (mtETC), a clinically validated antimalarial target. Our recent structure-activity relationship studies have identified key chemical elements underlying its potency and enabled the synthesis of derivative compounds with favourable antiplasmodial activity and enhanced selectivity for parasites over human cells. In vitro evolution experiments generated parasites carrying a single mutation in cytochrome b conferring high-level resistance to MMV1794211. Drug sensitivity profiling of the mutant revealed pan-strobilurin resistance; however, no cross-resistance was observed with established antimalarials or other mtETC inhibitors currently under investigation. Ongoing work includes assessing the fitness costs associated with strobilurin-resistance mutations and developing derivative compounds that remain active in resistant parasites. Collectively, our study provides a comprehensive evaluation of strobilurins as a chemically distinct class of antiplasmodial agents and highlights their promise for future therapeutic development.

Bovine Trichomonosis, caused by the protozoan parasite Tritrichomonas foetus, poses significant economic threats to the global cattle industry. In Australia, disease control relies heavily on accurate diagnosis followed by culling infected bulls. However, conventional diagnostic methods, including culture and quantitative PCR (qPCR), are limited by the complex microbial environment of the bull prepuce, which leads to false-negative and positive results. These arise from T. foetus low abundance and cross‑reactive trichomonad species.

To address these limitations, we developed and tested a diagnostic test using Oxford Nanopore Technologies (ONT) long‑read sequencing to simultaneously detect multiple venereal-transmitted organisms, T. foetus and Campylobacter fetus subsp. venerealis. Using artificially spiked DNA samples and field samples, our pipeline improved sensitivity and specificity, achieving detection limits for T. foetus down to 0.4 ng and identifying 50.0% more positives (n =9) than qPCR (n =6) across samples from persistently infected bulls. Simultaneously detecting C. fetus subsp. venerealis down to 0.04 ng, lower than its 0.1 ng qPCR limit.

These findings highlight the potential of ONT long-read sequencing as a robust, single-test alternative for accurately detecting T. foetus and C. fetus subsp. venerealis in bulls. Its adoption would enhance disease surveillance and help safeguard the Australian cattle industry.

Cyathostomin infections caused by small strongyle worms are among the most prevalent and clinically significant parasitic diseases affecting horses worldwide. Although considerable research has focused on identifying host–parasite interaction markers in serum and faecal samples, the salivary proteome remains largely unexplored. Saliva represents a non-invasive biofluid that may enable monitoring of infection status and immune responses, while providing access to both host and parasite derived proteins.

In this study, the salivary proteome of four naturally infected horses was analysed at two timepoints using mass spectrometry. Two preparation approaches were evaluated to determine the feasibility of saliva for parasite protein detection: direct digestion of neat saliva using S-Trap protocol, and on-bead enrichment method employing hydrophilic interaction liquid chromatography (HILIC) beads to concentrate polar and extracellular vesicle-associated proteins. Samples were analysed by data-independent acquisition (DIA) on an Orbitrap Astral mass spectrometer, searched in Spectronaut using DirectDIA, and analysed in the DIA Analyst platform.

Across all samples, 2,535 proteins (2,448 groups) were identified. Among these, 34 Cylicocyclus nassatus proteins were detected with more than one unique peptide, including 28 proteins with no detectable homology to the host proteome. These findings demonstrate the potential of equine saliva for detecting parasite and host immune proteins.

Gastrointestinal (GI) nematodes are a major constraint to dairy productivity and heifer growth globally and are of particular concern in subtropical Queensland. In this study, we collected fresh faecal samples from the paddock of 43 dairy farms, categorised by age into six groups: calves, weaners, heifers, springers, milking cows, and dry cows. Faecal egg counts (FEC) were performed using the modified McMaster technique, and species identification was conducted through larval culture, with morphological characterization of third-stage (L₃) larvae. All 43 farms had at least one age group positive for GI parasites, indicating widespread endemic infection throughout the region. The overall parasite prevalence was 78.7%, with infection burden varying by age: heifers had the highest FEC (mean EPG of 226.7; 26 to 553), followed by weaners (235.7; 31 to 651), and milking cows (75.8; 5 to 180). Calves were mostly uninfected. Egg counts did not differ significantly between farms (Kruskal–Wallis, p > 0.05) but varied strongly within age groups (H=107.5, p<0.001), with a clear age‑dependent gradient (p < 0.001). Haemonchus spp. (47.2%) and Cooperia spp. (43.1%) dominated, comprising ~90% of larvae, underscoring the need for regular, age‑based faecal monitoring to guide targeted parasite control on dairy farms in subtropical Queensland.

Strongyloides westeri is an intestinal threadworm of foals (≤16 weeks) that can cause dermatitis, diarrhoea and respiratory signs depending on parasite burden and transmission route. Despite its clinical relevance, epidemiological data on infections in Australian foal populations are limited. Additionally, traditional diagnostic methods such as faecal flotation techniques may have lower diagnostic sensitivity, highlighting the need for improved molecular diagnostic methods. This study aims to develop and validate a quantitative PCR (qPCR) assay for the detection and quantification of S. westeri in faecal samples from Australian foals. Following McMaster faecal egg count (FEC), DNA extracted from samples will be screened for Strongyloides spp. using a qPCR assay targeting the 18S ribosomal RNA region. The positive amplicons will be submitted for sanger DNA sequencing. Sequence analysis will be undertaken using Geneious Prime and alignments made with GenBank references, followed by species identity confirmation using phylogenetic analysis. Although experimental work is ongoing, it is anticipated that the qPCR assay will have greater sensitivity and accuracy compared to the traditional FEC. This will be the first study to provide molecular epidemiological data of S. westeri in Australian foals and support sustainable equine health management.

Ticks and tick‑borne diseases (TTBDs) are an emerging threat to human and animal health and livestock productivity. This study assessed knowledge, attitudes and practices (KAP) regarding TTBDs among cattle producers in Western Australia. A 44-item questionnaire (covering farm demographics, knowledge of tick-borne diseases, attitudes towards prevention and control, and on-farm management practices) was utilised to survey cattle producers in the region. The survey was distributed through regional biosecurity groups, field visits, and online platforms. Among 108 producers participated, 83 were included in the final analysis. Overall knowledge was low, no respondent (0/83) correctly answered all six knowledge questions, and the mean score was less than 1 out of 6. Most participants recognised the adverse effects of ticks on cattle health (81.3%), however, awareness of zoonotic risks (18.8%) and diseases such as bovine theileriosis (BATOG) (31.3%) was limited. Only 26.5% of the respondents reported favourable attitudes towards tick prevention and control. Management practices were variable, with 30.7% classified as poor, 47.6% as moderate, and 21.5% as optimal. Acaricides were commonly used (63.8%). This study reveals a critical low knowledge among cattle producers in WA and inconsistent control practices. Improved on-farm tick management including appropriate control methods, and disease awareness is required.

Gastrointestinal parasites are a significant health concern for horses, affecting their overall health and performance. Detecting intestinal parasites in horses is crucial for maintaining their health and preventing disease outbreaks, which can lead to significant economic and performance-related losses. This research provides essential data that can inform better management practices and parasite control strategies, ultimately enhancing horse health and productivity. This study was conducted to estimate the prevalence of different gastrointestinal parasites in horses in Riyadh, Saudi Arabia, and to investigate the relationship between infection rates and the horses' age, sex, and species. A total of 113 fecal samples from horses were gathered and examined using NaCl flotation and direct fecal smear techniques to detect gastrointestinal parasites. The results showed that among the 113 samples examined, 44 (38.93%) were found positive for various gastrointestinal parasites. The detected parasites included Entrobius spp. (10.6%), Eimeria spp. (6.19%), Anoplocephala spp. (2.65%), Parascaris equorum (1.76%), Ascaris spp. (1.76%), and Gastrodiscus spp. (0.88%). Additionally, 29.2% of the infections were attributed to unidentified oocysts, larvae, or eggs. These findings suggest that gastrointestinal parasites are common in horses in Riyadh, Saudi Arabia.

The treatment of malaria, an infectious disease caused by Plasmodium parasites, relies on numerous chemotherapies that are hampered by drug resistance. New antimalarial drugs with mechanisms of action different to currently used drugs are required to combat Plasmodium drug resistance. Robenidine is an antiparasitic drug with modest activity against P. falciparum (50% growth inhibitory concentration; IC₅₀ 0.76 µM) and unknown mode of action. To aid in the identification of new antimalarial drug candidates, a library of robenidine analogues was assessed for improved activity and selectivity for P. falciparum over mammalian cells. Multiple compounds with hit and early-lead activity and selectivity were identified, including NCL123 and NCL146, which demonstrated P. falciparum IC₅₀ values <0.1 µM and selectivity indices of >100. While in vivo studies with NCL123 and NCL146 demonstrated that further optimisation is needed to facilitate cures in mice infected with P. berghei ANKA, both compounds were well-tolerated and structural activity relationships have been useful in identifying avenues to improve this activity. Preliminary mode of action studies with NCL146 also suggest that this compound may have an unexploited mode of action associated with lipid biosynthesis, membrane function and cellular trafficking.

Proguanil is a used in combination with atovaquone to prevent and treat malaria. Activity of proguanil is thought to be due to its ability to potentiate atovaquone activity and the potent antiplasmodial activity of its in vivo metabolite, cycloguanil. Plasmodium falciparum resistance to cycloguanil is known to be due to mutations in the dihydrofolate reductase, but studies on proguanil resistance are lacking given it was essentially considered a prodrug. We overturned this dogma showing that proguanil has slow action activity against P. falciparum laboratory lines in vitro (Pf3D7 IC₅₀ 110 nM)¹. Here, we investigated thirteen culture-adapted P. falciparum isolates collected in eastern Uganda with low ex vivo sensitivity to proguanil². The in vitro sensitivity of the culture-adapted isolates to proguanil was assessed using 48 h, 72 h and 96 h growth-inhibition assays. While proguanil resistance was not confirmed in these studies, proguanil demonstrated slow-action activity against field isolates with 96 h IC₅₀ values of approximately 80-430 nM. These data extend our understanding of proguanil action to more clinically relevant field isolates, adding to findings that indicate that the antiplasmodial activity of the proguanil-atovaquone combination may be more complicated than previously thought.

Malaria remains a persistent global health threat leading to around 600,000 deaths annually. Resistance of Plasmodium parasites to most available drugs is significantly impacting prevention and control efforts. Combating resistance requires a clear understanding of drug mode of action, which remains incomplete for several current options. One such drug combination is atovaquone-proguanil. Atovaquone targets the cytochrome bc1 complex, and proguanil, originally developed as a prodrug, is converted in vivo to the dihydrofolate reductase (DHFR) inhibitor cycloguanil. While proguanil has been shown to potentiate the activity of atovaquone in vitro, for decades proguanil was thought to lack intrinsic activity. However, we showed that proguanil has a potent slow-acting in vitro antiplasmodial activity that is distinct from the folate metabolism pathway1. To try to identify the target/s of proguanil’s slow-action activity, we employed solvent-induced protein precipitation combined with mass spectrometry to assess protein stability in the presence of proguanil. Here, data will be presented on one of several candidate proteins stabilized by proguanil. Conditional knockdown is being performed in wildtype Plasmodium falciparum using TetR-DOZI and CRISPR-Cas9 Guide RNA constructs. Future work will include confirmation of knockdown and phenotypic assays with proguanil to assess changes in sensitivity to this drug.

The Australian sheep blowfly, Lucilia cuprina, is the principal cause of flystrike (cutaneous myiasis) in sheep, incurring losses of ~AU$320 million annually and inflicting severe welfare impacts in sheep. Current treatment includes insecticides and surgical mulesing; however, heavy reliance on insecticides has accelerated resistance, while mulesing draws strong public opposition due to animal welfare concerns—undermining both ethics and economic outcomes. Sustainable alternatives will be explored through the discovery and validation of novel blowfly-specific molecular targets identified using population genetic data from blowfly populations across Australia. RNA interference (RNAi) will be used to silence top molecular targets and protein modelling, and larval bioassays will be used for identifying and testing top candidate compounds. This project is expected to provide validated targets and candidate compounds for L. cuprina as the basis for novel larvicides to overcome existing insecticide resistance and reduce dependence on mulesing.

Gastrointestinal parasitic infections (GIP) are a major constraint to small ruminant production in Sri Lanka, yet a comprehensive synthesis of their epidemiology and anthelmintic resistance (AR) is lacking. This systematic review synthesised evidence on GIP, AR, and key knowledge gaps. A search of Web of Science, PubMed, Scopus, and CAB Abstracts (1971–January 2025), cross-checked with Google Scholar, identified 34 studies. Study quality was assessed using the Joanna Briggs Institute (JBI) prevalence checklist, and apparent prevalence was adjusted to true prevalence using the Rogan–Gladen estimator. In goats, prevalence ranged from 74–78%, while sheep showed seasonal prevalence of 84% (dry) and 92% (wet), with higher true prevalence after diagnostic adjustment. Predominant nematodes in goats were Haemonchus contortus (90%), Oesophagostomum columbianum (88%), Trichostrongylus colubriformis (76%), and Strongyloides spp. (72.5%). In sheep, Haemonchus contortus, Toxocara spp., and Trichuris spp. were common. AR was documented against benzimidazoles and levamisole, with Haemonchus spp. consistently implicated. Estimated economic losses in goats were LKR 230 million, based on limited data. Studies were heterogeneous in diagnostics, limiting comparability. These findings highlight critical gaps in parasite prevalence, species distribution, and AR, and underscore the need for coordinated, standardised national surveillance for sustainable parasite control in Sri Lanka.

Intestinal parasitic diseases remain a significant public health burden globally. There is an increasing need for a versatile, sensitive, and cost-effective mechanism for near-elimination and post-elimination surveillance systems for population-wide. Traditional faecal sampling from individuals across large populations is expensive, intrusive, and impractical. Wastewater-based epidemiology is a population-level surveillance tool.

A series of multiplex qPCRs was chosen to detect common parasites of public health importance. Limits of detection and specificity for human-infecting species were determined. Influent wastewater from an urban treatment plant was spiked with a known concentration of cysts, eggs, and larvae of the target parasites (Giardia duodenalis, Ascaris lumbricoides, Trichuris trichiura, Necator americanus) and processed using three concentration protocols. DNA was extracted using Powersoil Pro (Qiagen) kits, and four Taqman multiplex qPCRs were performed, including human DNA and internal amplification control targets.

The diagnostic qPCRs chosen demonstrated high sensitivity and specificity. Wastewater application of the multiplex was successful. Initial results indicate that centrifugation or sieve selection prior to DNA extraction was necessary to detect parasitic DNA in wastewater samples. Processing raw wastewater or wastewater sediments proved ineffective.

This work establishes a methodological basis for implementing wastewater-based population-level surveillance for soil-transmitted helminths and intestinal protozoa.

A 71-year-old man was found incidentally to have a crusted lesion on the floor of the nose during resection of a pituitary tumour. Microscopy showed an inflammatory infiltrate with ovoid cells, staining positive with PAS and silver raising concern for a free-living amoeba (FLA). However, multiplex PCR for Acanthamoeba species, Balamuthia mandrillaris and Naegleria fowleri was negative. There was no clinicoradiological evidence of meningoencephalitis. We reviewed the biology, clinical features and diagnosis of FLA infections.

Amongst 26 cases of rhinosinusitis, Acanthamoeba species were implicated in 25 and Entamoeba histolyticain one. The average age of patients was 43 mostly males. 11 patients died. Histopathology was used for diagnosis in 21 cases, PCR in 7. In two cases both were utilised. Culture was used in 1 case. All Acanthamoeba cases were associated with immunosuppression; HIV (17), CLL and solid organ transplant (3), haematopoietic stem cell transplant (1) and one hypogammaglobulinaemia.

Our asymptomatic case represents a dilemma, without literature to guide management in an incidental finding of amoeba in histopathological specimen especially after breach of the nasal mucosa for biopsy. FLA are ubiquitous and are found as carriers in the nose of volunteers. Disease likely affects immunosuppressed patients with compatible inoculation route.

Soil-transmitted helminths (STHs) cause some of the most prevalent neglected tropical diseases worldwide. Despite this, our understanding of diversity across hosts and extent of animal contributions to human infections remains limited. We screened 2,285 humans and 148 dogs from Vanuatu for STHs of public health importance using multiplex qPCR. A subset (n=148 humans, 148 dogs) were sequenced using a nanopore-based approach to characterise the ‘nemabiome’. Given evidence for zoonotic transmission of Ancylostoma ceylanicum and Strongyloides stercoralis, we also sequenced a large region of the cox1 gene in these species to assess haplotype diversity and infer zoonotic STH transmission dynamics. Overall, qPCR detected >1 STH in 48.5% of humans and 97.3% of dogs. Specifically, a prevalence of 8.9% and 64.9% were observed for A. ceylanicum and 3.0% and 47.3% for Strongyloides spp. in humans and dogs, respectively. Nemabiome analysis revealed a greater diversity of intestinal parasites than that detected by qPCR in both hosts. These findings indicate a high abundance of STHs across Vanuatu, supporting the need for mass-drug administration. Molecular data suggest canine-human transmission may be occurring, however further sequencing at nuclear and mitochondrial loci will resolve transmission dynamics and inform the need for control approaches that include animal reservoirs.

Introduction

Echinococcus granulosus sensu stricto (s.s.) is a zoonotic parasite that infects a wide spectrum of hosts. A substantial public health concern, this cestode causes socioeconomic burdens to resource-limited pastoral communities globally. The G1 and G3 genotypes of E. granulosus s.s. are associated with human and animal infection, with G1 predominating. However, an opposite trend has been observed in Pakistan, where the G3 genotype is more common.

Materials and Methods

In the present study, genomic DNA was extracted from hydatid cysts obtained from cattle in Multan, Pakistan and processed for whole-genome sequencing (Illumina NovaSeq). Complete mitochondrial genomes were assembled and population genetic analysis was performed and compared to those from public databases.

Results

The results indicated that the G3 genotype of E. granulosus s.s. is highly prevalent among the cattle of Pakistan. Using whole mitogenome data (13,610 bp), when compared to the G1 reference sequence, G3 exhibited substantial variation at 48 diagnostic positions, indicating a distinct mitochondrial lineage. Genetic variability indices revealed high haplotype diversity and low nucleotide diversity.

Conclusions

This study reports the first complete mitogenome of the G3 genotype of E. granulosus s.s. from Pakistan, providing a foundation for future genetic analyses from other regions of the world.

Ascaris spp. infect approximately 804 million people, causing substantial malnutrition, stunting and morbidity in school-aged children in low-income populations, exceeding 750,000 Disability Adjusted Life-Years annually. Current single-dose oral anthelmintics offer no sustained protection nor interruption of transmission. Interventions that prevent infection are needed. During early infection, Ascaris larvae migrate from the host intestine to the liver and lung before maturation in the small intestine. This process is thought to be passively directed by host blood flow; however, multiple studies suggest active chemotaxis. Here, we use larval agar migration assays and microfluidic chemotaxis arenas to demonstrate that freshly-hatched Ascaris suum larvae exhibit liver-specific chemotaxis in vitro, including increased migration distance, speed, and directional movement toward liver gradients, and reduced turning behaviours, relative to lung or RPMI controls. These responses coincided with specific transcriptional up-regulation of chemotaxis receptors, signalling markers and metabolic pathways in liver relative to lung or RPMI. Our findings provide strong evidence of organ-specific, active chemotaxis in Ascaris infection, with implications for other major human helminthiases, and identify conserved chemosensory pathways as promising therapeutic targets. Disrupting chemosensory-guided navigation could prevent larval migration, offering a novel strategy to combat ascariasis.

RNA-binding proteins (RBPs) regulate splicing, RNA silencing, and translational repression in eukaryotes. Many are conserved from yeast to humans but are absent or rudimentary in prokaryotes, suggesting an early emergence of “eukaryotic-innovative” RBPs. We hypothesised that these RBPs arose long before yeast and are retained in Giardia, an early-diverging eukaryote. To test this, we built a phylogenomic atlas of RBP families across the tree of life and analysed domain topology, domain co-occurrence, and intrinsically disordered regions (IDRs) to define their architectural evolution. We then characterised the Giardia RBPome using domain- and structure-informed annotation integrated with transcriptomics, proteomics, and RNA–protein interactome capture to identify canonical and non-canonical (“moonlighting”) RBPs. Direct RNA targets and regulatory networks of representative eukaryotic-innovative RBPs, including PUF, DDX3X, EIF4A, and PGK, were resolved using enhanced CLIP-seq and RBP immunoprecipitation. Functional significance was assessed by CRISPRi-based genetics, and condensate behaviour was tested using phase-separation assays in vitro and in vivo. Our analyses show that multiple eukaryotic-innovative RBPs are already present in Giardia, with simplified but functional architectures, conserved RNA–protein networks, regulatory phenotypes, and condensate-like behaviour, establishing Giardia as a minimal model for the earliest evolution of eukaryotic post-transcriptional control.

Giardia duodenalis is a gastrointestinal parasite causing ~200 million symptomatic infections annually, disproportionately in lower socioeconomic tiers and children. Chemotherapeutic interventions are limited to nitroheterocyclic antibiotics such as metronidazole. However, high doses are toxic and drug-resistant treatment failures occur in up to 20% of cases, highlighting the urgency of novel and safer chemotherapeutics. Here, we target the disproportionate kinome of G. duodenalis with small-molecule inhibitors to reveal novel antigiardials and their molecular targets for next generation antiparasitics.

Using "Click" chemistry, we immobilised a potent drug-like kinase inhibitor to azide-agarose supports and identified a high-affinity kinase domain-containing protein (GiK5) as the putative target for this inhibitor. We validated recombinant GiK5-inhibitor engagement through differential scanning fluorimetry, native mass spectrometry and the fluorescent ADP-glo assay. Further, we conducted a multiplexed CRISPR-interference knockdown which showed lower GiK5 abundance at the protein level contributed to slower parasite growth, suggesting the likely essentiality of this protein in the parasite.

We reveal this likely druggable kinase in G. duodenalis and this workflow incentivises high-throughput, target-centric screening campaigns for structure-guided drug-discovery, as well as repurposing clinically-approved kinase inhibitors for chemotherapeutic interventions against this parasite.

Programmed cell death (apoptosis) is a fundamental process in metazoans, extensively characterised in vertebrates but comparatively understudied in invertebrates beyond the model organisms Caenorhabditis elegans and Drosophila melanogaster. Here, we present the first reconstruction of the intrinsic apoptosis pathway in the parasitic nematode Haemonchus contortus, a blood-feeding pathogen of ruminants responsible for substantial global production losses. Using C. elegans proteins as a reference, we integrated genome-wide homology searches with structural modelling and developmental transcriptomic and proteomic analyses to identify apoptosis regulators in H. contortus. Homologues of all canonical pathway components were identified, including CEP-1, EGL-1, CED-9, CED-4 and CED-3, together with modulators such as DRE-1 and PUF-8. Structural analyses revealed conservation of key interaction complexes (CED-9:CED-4 and CED-4:CED-3), whereas EGL-1 and CEP-1 retained critical structural domains despite marked sequence divergence. Transcriptomic profiling showed that Hc-ced-9 and Hc-ced-3 are constitutively expressed across developmental stages, whereas Hc-cep-1 and Hc-egl-1 display stage-specific transcription. Proteomic data confirmed the presence of Hc-CED-9, Hc-CED-4 and Hc-CED-3 in at least one life stage but did not detect Hc-EGL-1 or Hc-DRE-1. Discordances between transcript and protein abundance, particularly for Hc-EGL-1, suggest post-transcriptional regulation. Future efforts are needed to elucidate the apoptosis pathway across members of the phylum Nematoda.

Intrinsic apoptosis is a form of programmed cell death that governs development, tissue homeostasis and stress responses in animals. In nematodes, the pathway was first genetically defined in the free-living nematode Caenorhabditis elegans, yet how it has diversified and operates across the phylum, which encompasses parasites of humans and animals spanning clades I–V, remains poorly resolved. Here, we synthesise genomic, structural and functional data to establish a framework for intrinsic apoptosis in nematodes. Although the core CED-9-CED-4-CED-3 module is retained, its developmental deployment and regulatory processes remain largely uncharacterised beyond C. elegans. Unlike the corresponding BCL-2-regulated apoptotic pathway in vertebrates, C. elegans lacks a canonical BAX/BAK-driven mitochondrial permeabilisation system, and whether comparable mitochondrial amplification mechanisms operate in other nematodes remains unclear, particularly in species encoding multiple CED-9/BCL-2-like proteins. This alternative regulatory configuration, coupled with structural divergence of nematode CED-9 and CED-4-like proteins from their vertebrate orthologues, suggests a distinctive evolutionary trajectory for apoptotic regulation in nematodes. By integrating biological with emerging structural insights, we define a foundation for studying apoptosis across clades I–V and assess its potential for anthelmintic target discovery.

Schistosomiasis-induced fibrosis, driven by host immune response to trapped eggs, is the principal cause of pathology and schistosomiasis-related morbidity. While praziquantel effectively clears adult parasites, no therapies exist to target egg or egg-induced tissue fibrosis. To address gaps in fibrosis-related cellular mechanisms and identify novel antifibrotic targets, we used single-cell spatial transcriptomics with a 5000-gene mouse panel to map the molecular architecture of hepatic and intestinal fibrotic regions in Schistosoma mansoni-infected mice. Results showed that schistosome-induced granulomas are highly organized and overlap with fibrotic regions in both tissues at 8 weeks post-infection. Differential expression analysis identified 1175 genes in 534k liver cells and 807 genes in 259k intestine cells significantly altered between healthy control and infected samples. These DEGs are strongly associated with cytokine and interleukin signaling, TGF‑β pathway, and extracellular matrix organization. The cell types enriched in infected regions included macrophages, collagen-producing cells (liver-activated hepatic stellate cells, intestine-activated fibroblasts), eosinophil-like cells, T, B, and plasma cells. Furthermore, cell-cell interaction analysis revealed strong interactions between macrophages and collagen-producing cells in infected tissues. While many identified ligand-receptor pairs are organ-specific, a core signature of 5 pairs (including Col1a2/Tgm2-Itgb1) was shared across tissues, representing potential pan-tissue therapeutic targets for schistosome-induced fibrosis.

Alternative splicing is a major mediator of eukaryotic gene expression, operating co-transcriptionally and post-transcriptionally to influence cellular function, development, and differentiation. During alternative splicing, trans-acting factors interact with cis-elements within pre-mRNA transcripts to produce divergent proteoforms. In eukaryotes, mosttrans-acting factors work in conjunction as the spliceosome, a dynamic ribonucleoprotein complex comprised of catalytic RNAs and hundreds of proteins. Previous studies sought to comprehend the intricate mechanisms of the spliceosome within a simplistic system, utilising the model organism Saccharomyces cerevisiae. However, there exists a eukaryote more basal than yeast – Giardia duodenalis, an intron-poor enteric parasite, evolved over 500 million years earlier. Our bioinformatic analyses indicated that splicing proteins in Giardia are minimal, both in number and structure. This raises the question: In deeply-branching eukaryotes such as Giardia, does splicing occur spliceosomally or via a more primordial method? To understand the system of splicing in Giardia, we conducted an in vitro splicing assay involving both conventional and chimeric intron-containing transcripts. In parallel, we established a nuclear proteome to locate and validate the suite of Giardia splicing proteins. By implementing a multiomic approach, our study offers a glimpse into the origins and evolution of alternative splicing following the inception of eukaryotic life.

Mosquitoes are the most significant vectors of human and animal diseases. Worldwide, there are over 3,500 extant species, each with varying ecological behaviours. Consequently, mosquito populations are dynamic, shifting over time and space. As a result, the risk of disease transmission is also dynamic, as it relies on the prevalence and abundance of known competent vectors. Therefore, understanding how mosquito populations shift is essential to accurately assess mosquito-borne disease transmission risk.

To investigate spatiotemporal effects on mosquito populations, 4,469 mosquitoes (representing 15 species) were collected from 10 locations within the Perth metropolitan area between 2022 and 2024. Climate variables (rainfall and temperature) and landscape coverage (within 2.5 km) were obtained for each location and trapping event. Generalised linear models determined the influence of spatiotemporal variables on mosquito population diversity indices (Hill’s numbers) and species abundance.

Mosquito population diversity was not significantly influenced by landscape coverage; instead, diversity was driven by climate variables. Mosquito species exhibited diverse temperature and rainfall preferences, indicating that competent vectors for endemic mosquito-borne diseases are prevalent year-round in Perth. Our findings underscore the importance of integrating climate-driven and land-use data into mosquito surveillance and control programs to improve predictions of mosquito-borne disease transmission risk.

Cuora flavomarginata, an endemic terrestrial turtle in Asia and a Class I protected species in Taiwan, remains understudied in terms of parasite diversity and zoonotic risk. This study investigated endo- and ectoparasites, with a One Health focus on tick-borne pathogens. Samples were collected from 12 turtles in Shizi Township, yielding 154 ticks and 6 fecal samples. All ticks were identified as Amblyomma geoemydae using morphological and molecular approaches. A total of 51 tick DNA samples were screened by PCR for Francisella spp., Rickettsia spp., Coxiella burnetii, and Ehrlichia spp., while fecal samples were examined for endoparasites. Francisella spp. showed a notably high detection rate (86.3%), followed by Rickettsia spp. (27.5%) and C. burnetii (2.0%); Ehrlichia spp. were not detected, and no endoparasites were found. The high prevalence of Francisella spp. suggests that reptile-associated ticks may contribute to its environmental maintenance. Given the unclear transmission pathways of tularemia in Taiwan, these findings highlight a previously underrecognized component in its ecology. This study underscores the importance of integrating wildlife, vector, and pathogen surveillance to better understand zoonotic disease risks within a One Health framework.

Bats are key ecological species and recognised reservoirs of diverse pathogens, yet their role in tick–host–pathogen systems in Australasia remains poorly understood. This study systematically synthesises current knowledge on bat-associated ticks and tick-borne pathogens (TBPs) and identifies critical gaps. Following PRISMA guidelines, literature was retrieved from major databases, including studies reporting ticks on bats or within their environments, and those investigating TBPs. Risk of bias was assessed using an AXIS-based framework. Of 512 studies screened, 35 met the inclusion criteria. Fourteen typical and nine incidental tick species were reported from bats, predominantly in Australia and New Guinea. Only one bacterial pathogen, Rickettsia japonica, was detected in the typical bat-associated tick Argas dewae. The non-typical tick Ixodes holocyclus was associated with paralysis in bats, while A. dewae may infest other hosts, including humans. Most studies relied on opportunistic sampling and morphological identification, with minimal use of molecular tools and limited investigation of pathogen diversity or bat health impacts. These findings highlight major gaps in surveillance, molecular characterisation, and pathogen detection, underscoring the need for integrated One Health approaches to better understand bat–tick–pathogen dynamics in Australasia.

Antibody Fc regions have important roles in clearance of Plasmodium falciparum infected erythrocytes. One such role is engagement with Fcg receptors on host leukocytes. In the case of FcgRIIIa and b, this interaction is greatly enhanced when the IgG glycan is afucosylated. In this study, we used a fucose-sensitive enzyme-linked immunosorbent assay, FEASI, to assess afucosylation in IgG specific for placental malaria protein VAR2CSA from n=139 malaria-exposed pregnant Malawian women, correlating these data with mass spectrometry-based analysis. Furthermore, we measured the effect of afucosylation on Fc-mediated leukocyte functions, using both plasma and a monoclonal antibody, PAM2.8, with varying levels of afucosylation.

Results showed significantly higher levels of VAR2CSA-specific IgG afucosylation in women with no placental malaria measured by FEASI (p < 0.0001), which correlated strongly with mass spectrometry analysis (R = 0.8, p < 0.0001). In addition, highly afucosylated IgG mediated significantly greater neutrophil phagocytosis of antigen coated beads and induction of NK cell degranulation by IEs.

Afucosylated IgG to VAR2CSA, measured by FEASI or mass spectrometry, was a correlate of protection from placental malaria, and afucosylated IgG activated NK cells and neutrophils. Naturally acquired or therapeutic afucosylated IgG antibody could have a role in protection from malaria infection.

Helminth infections, whether experimental or naturally acquired, are increasingly recognised as potent modulators of human immunity, with protective effects across a range of inflammatory diseases. Much of this activity is driven by excretory/secretory proteins (ESPs), complex mixtures of bioactive molecules that act at the host–parasite interface to reshape immune responses and suppress inflammation. Despite this, the therapeutic use of live helminths remains limited due to safety concerns, complex life cycles, and variable host responses. Consequently, focus has shifted toward isolating individual ESPs as more tractable, drug-like candidates. To address this, we generated a recombinant library spanning the secretomes of both larval and adult stages of the human hookworm Necator americanus. This enabled systematic screening across in vitro and in vivo assays to identify proteins with immunoregulatory activity. To date, two distinct protease inhibitors have emerged, each displaying pronounced anti-inflammatory effects. Their independent identification via separate screening strategies underscores the library's versatility as a discovery platform. Their protease inhibitory activity has been confirmed in vitro, consistent with established roles for helminth-derived inhibitors in modulating host inflammatory pathways. Work is now focused on defining their mechanisms of action in vivo and assessing their potential as pre-clinical therapeutic candidates.

Effective malaria interventions are essential to reduce disease burden in children. Combining RTS,S vaccination with seasonal antimalarial chemoprevention was shown to enhance efficacy against clinical malaria among children by ~72% over the first year compared to either intervention alone. However, over four years, efficacy of this combination steadily decreased despite annual boosters. Protective antibodies were shown to peak following primary vaccination but became progressively lower with each annual booster. It is unknown what drives poor antibody responses to boosters, which likely vary between different antibody isotypes and antigenic targets, and if host factors such as viral co-infections and micronutrient deficiencies are implicated in this suboptimal response.

To address these knowledge gaps, we evaluated plasma samples from young children (n=1,929) in West Africa who received RTS,S and subsequent boosters with or without seasonal chemoprevention over four years as part of a phase-III clinical trial. We found that repeated vaccine doses had differential effects on antibody responses, which varied by the antigenic region of the vaccine. We also investigated the impact of host factors on these responses. By improving our understanding of the immune response to repeated booster doses, this work informs optimal RTS,S implementation strategies to improve vaccine efficacy and longevity.

The significant global burden of malaria, caused by Plasmodium spp., warrants novel treatment and prevention strategies. Plasmodium falciparum merozoite surface protein 2 (PfMSP2) has been a target of blood stage malaria vaccines, which have reached phase I/2b clinical trials. Strain-specific polymorphisms complicate the development of this antigen as a vaccine candidate; most PfMSP2 genes can be classified into two main alleles, FC27 and 3D7. Bioinformatic comparison indicates that MSP2 of Laverania species maintain conserved and intrinsically disordered property regions, and tend to have repeat structures that most closely resemble P. falciparum 3D7-like alleles. We successfully knocked out PfMSP2 in both FC27- and 3D7-like parasites, which did not influence blood stage growth but did increase potency of antibodies targeting another vaccine candidate, PfAMA1. The ama1 genotypes of parasites with 3D7- and FC27-like msp2 do not significantly differ, indicating the dimorphism of PfMSP2 is not significantly influenced by PfAMA1 diversity. Ongoing work is applying microscopy and gene editing techniques to understand the role of PfMSP2 in potentiating antibodies to AMA1. This work will define the impact of msp2 diversity on antibodies targeting other antigens and will help inform merozoite vaccine development.

Cryptosporidium and Giardia are major causes of gastrointestinal illness globally. In Australia, cryptosporidiosis is notifiable, yet molecular characterisation is not routinely performed, limiting detection of outbreaks and zoonotic transmission. In 2024, Australia recorded a 273% increase in cryptosporidiosis notifications, the third-highest rise globally. Here, we conducted an epidemiological investigation of Cryptosporidium and Giardia in faecal samples from patients with gastroenteritis in Victoria between 2020 and 2024. Samples underwent SSU and gp60 sequencing for Cryptosporidium and tpi sequencing for Giardia, and parasite load was estimated. Of 2,330 samples, 225 were Cryptosporidium-positive and nine Giardia-positive. One Giardia isolate was sub-assemblage AI, two AII, and six assemblage B. Seven species and 24 subtypes were identified, including eight novel subtypes. Cryptosporidium hominis predominated (85%), followed by C. parvum and C. meleagridis. Six C. hominis subtypes were detected, three linked to 11 recreational water-associated outbreaks in 2024; IaA12R3 and IeA11G3T3 were most frequent, with concordance analysis suggesting 52 additional cases. Multiple C. parvum subtypes were identified, including two linked to childcare and camp outbreaks, and novel human-infective subtypes of C. occultus, C. fayeri, and C. meleagridis were detected. These findings highlight Cryptosporidium diversity in Victoria and the value of molecular surveillance for public health.

Ixodes holocyclus (the Australian eastern paralysis tick) is a medically and veterinary important ectoparasite that produces potent neurotoxins, holocyclotoxins, causing rapidly ascending flaccid paralysis in companion animals, livestock and humans, often fatally. Despite its importance, the molecular basis of toxin production, host specificity and survival remains poorly understood because genomic and transcriptomic resources are limited. We generated the first chromosomal-scale genome for I. holocyclus using Oxford Nanopore long reads, Illumina short reads and Hi-C, and annotated genes with a hybrid de novo transcriptome, resolving alternative splicing with long- and short-read alignments. We scanned UTRs and upstream regions of complete genes for conserved regulatory motifs, including putative promoters. Comparative genomics, including synteny and phylogenomic placement, was performed, and ticks from 32 eastern Australian sites were sequenced to examine genomic diversity and its links to ecological adaptation and vector capacity. The 1.9 Gb assembly contains 13 chromosome-level scaffolds, 66% repetitive elements and 93.3% BUSCO completeness. Annotation identified a high-confidence gene set including protein-coding genes. Synteny with I. scapularis and I. ricinus revealed conserved supergene blocks. Together, these resources advance tick biology and support targeted control strategies against tick-borne diseases.

The European castor bean tick, Ixodes ricinus, is a major blood-feeding ectoparasite and vector of Lyme disease, tick-borne encephalitis and babesiosis. Reducing tick-borne disease requires an improved understanding of tick molecular biology across tissues and feeding stages. We applied long-read, full-length mRNA sequencing to refine gene models in the published I. ricinus genome and to profile salivary glands and ovaries across blood-feeding phases. Curated functional annotation was used to interpret pathways central to ectoparasitism. We defined 30,454 gene models, including 28,515 protein-coding genes. Functional curation provided insight into chemosensation, hematophagy, immune tolerance, reproduction and fecundity. Salivary glands showed feeding-associated metabolic rewiring, vesicle biogenesis, and secretome remodelling, consistent with anti-clotting, vasodilatory, anti-inflammatory, and immunomodulatory functions that counter host defences. Ovarian transcriptomes revealed early reprogramming of cell-fate pathways, cytoskeletal organisation, extracellular matrix dynamics, and immune regulation, consistent with priming for fertilisation and embryogenesis. This transcriptomic blueprint provides a high-resolution annotation resource for the I. ricinus genome and delineates tissue- and stage-specific programmes that underpin feeding and reproduction, providing a platform for mechanistic studies and improved tick control strategies.

Haemaphysalis longicornis is a parthenogenetic three-host tick of increasing veterinary and public health importance and is the principal vector of Theileria orientalis in Australasia. Experimental research on this species has been constrained by reliance on live vertebrate hosts, limiting scalability, standardisation and ethical feasibility. This study established a reproducible, host-free in vitro membrane feeding system for adult and nymphal stages. Ticks were fed using a silicone membrane-based system under controlled laboratory conditions. Field-collected adult females were evaluated across six independent experiments for attachment, engorgement, oviposition, egg hatchability and bloodmeal-to-egg conversion efficiency. Nymphal feeding performance was assessed across five experiments. Adult feeding was robust, with 67% (35/52) attachment and 74.3% (26/35) engorgement. Engorged females reached a mean weight of 161 mg and produced a mean egg mass of 67 mg (40% conversion efficiency), with >92% hatchability. Nymphs showed consistently high performance, with 88.4% (76/86) attachment, 97.4% (74/76) engorgement and 90.5% (67/74) moulting success. Engorgement typically occurred within 3–5 days post-attachment, and moulting within 2–4 weeks post-detachment. This system enables controlled, ethical and scalable experimentation and provides a powerful platform for studies of tick physiology, vector competence, acaricide screening and pathogen–vector interactions.

Ticks are obligate haematophagous ectoparasites, and species within the Rhipicephalus microplus complex are responsible for substantial economic losses to the cattle industry. Traditionally, laboratory rearing of ticks has relied on live animal hosts; however, increasing animal welfare concerns have driven the development of artificial tick feeding systems (ATFS). Here, we report the first successful consecutive artificial feeding of all life stages of the Australian cattle tick, Rhipicephalus australis, using a silicone membrane-based system. Success was achieved through optimisation of membrane thickness, incorporation of olfactory stimuli and strict contamination control, enabling continuous in vitro feeding of this one-host tick species. Using larvae aged 2-13.5 weeks, all developmental stages demonstrated good attachment and engorgement rates. This study represents the first demonstration of consecutive feeding of all life stages of R. australis without the use of live animal hosts. The developed ATFS provides a good platform for further investigating tick biology and tick–pathogen interactions under controlled conditions, as well as for evaluating acaricides/vaccine candidates. However, the limitations of this system include reduced oviposition and increased mortality indicating the need for further optimisation. Overall, this system supports the principles of the 3Rs (Replacement, Reduction and Refinement) in tick and tick-borne disease research.

Ticks are among the most important vectors of pathogens affecting livestock. Rhipicephalus australis (the Australian cattle tick) transmits several economically significant pathogens, including Anaplasma and Babesia spp. Increasing evidence suggests that the tick microbiome influences pathogen acquisition, persistence and transmission, thereby shaping vectorial capacity. This study characterised bacterial communities across larval, nymphal and adult stages of R. australis collected from cattle farms in Queensland, Australia. Following surface decontamination and DNA extraction, 16S rRNA gene sequencing was performed using the Illumina NextSeq 1000 platform, with downstream analyses conducted in QIIME 2 and R. A diverse bacterial community was identified, including Arsenophonus, Acinetobacter, Brevibacterium, Coxiella, Corynebacterium, Serratia, Stenotrophomonas, Escherichia–Shigella and Staphylococcus. Several taxa were consistently detected across all life stages, suggesting conserved or potentially symbiotic associations while others were stage-specific, indicating dynamic shifts in microbial community composition during tick development. Microbial network analyses further revealed distinct, stage-specific interaction patterns. These findings provide the first comprehensive insights into life stage-associated microbiome variation in R. australis and highlight the potential role of microbial communities in tick biology and pathogen transmission. This work establishes a foundation for microbiome-informed strategies to improve Australian cattle tick control and enhance livestock health and productivity.

Ticks are important vectors of pathogens affecting both humans and animals. For example, cattle ticks transmit Anaplasma and Babesia spp., causing substantial economic losses to the global cattle industry. Artificial tick feeding systems (ATFS) are increasingly used as alternatives to animal-based feeding to study tick biology, pathogen transmission and control strategies. However, their impact on the tick microbiome remains poorly understood. This study investigated the microbiome and microbial co-occurrence networks of the Australian cattle tick (Rhipicephalus australis) reared under ATFS conditions to establish baseline insights. Ticks were collected across multiple life stages, from larvae to engorged females, surface decontaminated, and processed for DNA extraction. Microbial profiling was conducted using 16S rRNA gene sequencing on the Illumina NextSeq 1000 platform, with downstream analyses performed in QIIME 2 and R. Artificially fed ticks harboured diverse microbial communities, with variation observed across life stages. Network analyses revealed distinct microbial interaction patterns, suggesting structured community dynamics under in vitro conditions. These findings indicate that ATFS environments, including the blood meal, membrane system, and absence of host-derived immune factors, may influence microbial community composition. Such effects should be considered when interpreting biological processes and vector competence in artificially fed ticks.

Theileria orientalis Ikeda has been a significant cause of disease in the Australian cattle industry for over a decade. To investigate how widespread T. orientalis Ikeda is across Australia, particularly in regions in and outside of known ranges of the vectors and in the absence of clinical disease, we conducted testing on herds of homebred adult cattle in New South Wales and Queensland. Molecular testing of 526 blood samples from 49 properties was performed to detect and quantify different T. orientalis genotypes present within these herds. Previous studies have shown that in clinical cases of theileriosis, the Ikeda genotype is detected in 88% of samples.

Preliminary findings suggest that T. orientalis Ikeda may be more widespread than previously thought, including in areas outside the known ranges of the tick vectors. These detections in herds that have not experienced clinical disease indicate that presence of the organism may be a necessary but not sufficient cause of disease on its own. These results help build a clearer understanding of clinical theileriosis in Australia and highlight the value of ongoing active surveillance. Future work to include additional Australian states is planned.

Cryptosporidium is a major enteric zoonotic protozoan parasite with a wide host range, causing gastrointestinal disease in humans and animals. Livestock, particularly calves, serve as an important zoonotic reservoir for Cryptosporidium parvum, the main zoonotic species. In the present study, 300 cattle faecal samples (200 calves and 100 adult cattle) from Western Australia (WA) were screened for Cryptosporidium by PCR and sequence analysis at the18S rRNA locus. Cryptosporidium parvum positives were analysed using multi locus sequence typing (MLST) of five polymorphic loci including the hypervariable gp60 gene and compared to previously typed human C. parvum samples from WA.

Results revealed that C. parvum was the only species detected in both calves and adult cattle with three C. parvum gp60 subtype families identified; IIaA18G3R1, IIaA19G4R1 and IIaA20G3R1. MLST analysis demonstrated limited genetic diversity among cattle and human C. parvum isolates, with most isolates clustering within a single dominant clade. The intermixing of host-derived isolates supports potential zoonotic transmission of C. parvum.

Cryptosporidium and Giardia are globally important enteric protozoa with broad host ranges and significant One Health implications. However, molecular epidemiological data in Australian goats remain limited. This study investigated the molecular prevalence, species composition and genotype diversity of Cryptosporidium and Giardia in Australian dairy goats. A cross-sectional survey (2023–2024) was conducted involving 386 goats (4 weeks–12 months old) from 61 dairy herds across six states. DNA samples were analysed using nested PCR and Sanger sequencing for species and assemblage identification. Cryptosporidium was detected in 6.5% of samples and Giardia in 9.3%. Three Cryptosporidium species were identified: Cryptosporidium xiaoi, C. ubiquitum, and C. muris, with the latter two recognised as zoonotic. Giardia duodenalis assemblages AI and E were detected, including five novel assemblage E variants. C. muris and a zoonotic Giardia sub-assemblage AI are reported for the first time in Australian goats, alongside two novel C. xiaoi genotypes. Although overall prevalence was low, the detection of zoonotic genotypes highlights the role of young goats as reservoirs for infection. The absence of C. parvum, a major zoonotic species globally, suggests distinct regional transmission dynamics. These findings provide baseline molecular data to inform surveillance, risk assessment and One Health management strategies in Australia.

Cryptosporidium spp. and Giardia duodenalis are globally significant enteric protozoa affecting livestock health, productivity and public health through their zoonotic potential. In pigs, these infections can impair growth performance and welfare, yet molecular epidemiological data in Australian production systems remain limited. This study aimed to determine the prevalence, distribution and genetic diversity of Cryptosporidium spp. and G. duodenalis in pigs across Victoria. A cross-sectional survey was conducted using 626 faecal samples collected from pigs of different age groups across 69 commercial farms. Samples were screened using nested PCR, and amplicons were subjected to Sanger sequencing. Phylogenetic analyses were conducted to identify species, assemblages/subtypes and determine their genetic relationships. Preliminary findings have identified Cryptosporidium scrofarum, C. suis and the zoonotic species C. ubiquitum, as well as G. duodenalis assemblages A and E. These results indicate the presence of both host-adapted and zoonotic genotypes in Victorian pig populations. This study will generate the first comprehensive molecular epidemiological dataset for these protozoa in Victorian pigs, providing critical insights into infection dynamics and zoonotic risk. The findings will inform evidence-based parasite control, strengthen biosecurity strategies and support sustainable productivity in the Australian pork industry.

Building on our previous report of high prevalence of soil-transmitted helminth (STH) infections among Myanmar schoolchildren (Aung et al., Infectious Diseases of Poverty, 2022), we conducted additional molecular screening of archival stool samples from the same cohort in Phyu Township, Bago Region, to investigate additional helminth infections. We also report finding of other helminths by Kato-Katz in the previous study that were not previously published. Stool samples utilised in this study were collected in 2016 and the DNA extracted in 2017 and kept stored at -20°C until further molecular characterisation in this study in 2025. Using quantitative PCR (qPCR), we detected Schistosoma DNA in two of 264 samples, Strongyloides stercoralis DNA in twelve, and Ancylostoma ceylanicum in eleven. Although sequencing of the Schistosoma-positive samples was unsuccessful, the molecular evidence aligns with other recent reports suggesting emerging or cryptic transmission of schistosomiasis in Myanmar. The epidemiology of schistosomiasis in the region remains poorly defined, highlighting the need for targeted snail surveys, environmental DNA (eDNA) monitoring, and host sampling to confirm transmission foci. This study demonstrates the added value of molecular diagnostics for complementing traditional parasitological methods and guiding surveillance and control strategies in areas of emerging endemicity.

Parasitic nematodes of the family Onchocercidae have co-evolved with vertebrate hosts for millions of years. Although morphology has traditionally underpinned species identification, many taxa are cryptic and difficult to distinguish, limiting accurate diagnosis. Clinically important genera include Dirofilaria, which causes heartworm disease in dogs and occasional zoonotic infections in humans. However, substantial gaps remain in our understanding of other Dirofilaria species and genotypes and their impacts on host health. This study aimed to generate a comprehensive morphological and molecular resource for a newly identified Dirofilaria species. Using long-read PacBio and Hi-C sequencing, we assembled and characterised both mitochondrial and nuclear genomes. The nuclear genome comprises four autosomes and one sex-linked scaffold, encoding 9,658 genes. Comparative analyses with related filarial nematodes revealed conserved chromosomal structure alongside lineage-specific rearrangements. We identified 881 predicted excretory/secretory proteins enriched in immune-related pathways such as proteolysis, lysosomal function and antigen presentation. Notably, 26% of these proteins were unique, many associated with host–parasite interactions, immune evasion and metabolic adaptation. This genome fills a key gap in filarial resources and supports advances in epidemiology, host adaptation studies and diagnostic development.

Understanding how biological processes and interactions unfold within intact organisms requires approaches that link anatomical organisation with spatially resolved molecular information. Although spatially integrated analyses have transformed vertebrate biology, comparable frameworks remain limited for many non-model invertebrates despite their ecological and biomedical importance. Freshwater snails represent a particularly informative system, functioning both as key components of aquatic ecosystems and as intermediate hosts for numerous parasitic organisms. Here we establish an integrated spatial–molecular analytical framework for investigating biological processes within a lymnaeid snail using infection with the liver fluke Fasciola hepatica as a model host–parasite system. The approach combines whole-organism three-dimensional imaging using X-ray microcomputed tomography with serial histopathology, bulk RNA sequencing and spatial transcriptomic analysis. Integrating these complementary datasets enables parasite distribution to be examined within the anatomical structure of the host while simultaneously linking tissue pathology with spatially resolved gene transcription patterns. Beyond the present host–parasite system, this framework provides a foundation for investigating a broad range of biological processes in freshwater snails, including environmental responses, neurobiology and host–parasite interactions.

Understanding the population structure of Simulium damnosum, the blackfly vector of Onchocerca volvulus, is fundamental to predicting onchocerciasis transmission risk and optimising intervention strategies. We investigated the genetic structure of S. damnosum. S. damnosum specimens were collected from 24 localities across Ghana via human landing catches. Following genomic DNA extraction and short read-sequencing, genome-wide nuclear SNPs were generated and analysed using DAPC and ADMIXTURE to delineate ancestry clusters. Five genetically distinct clusters were identified, broadly corresponding to Ghana's major agro-ecological zones: two savannah clusters (GS/SS-A and GS/SS-B), a Coastal Savannah (CS), and two Semi-Deciduous Forest clusters (SDF-W and SDF-E). The unexpected subdivision of the SDF ecozone into two divergent lineages, with the Bosomase population forming a genetically isolated western unit, suggests that O.volvulus transmission dynamics may differ markedly across this zone despite shared ecological classification. Populations at ecozone boundaries showed elevated admixture (gene flow), indicating contact zones where vector populations and parasite strains may intermix. These results indicate that S. damnosum populations in Ghana should not be treated as a single panmictic unit for epidemiological modelling or parasitological surveillance. Parasite-vecto compatibility studies stratified by genetic cluster are warranted to fully characterise transmission risk across Ghana's heterogeneous landscape.

Strongyloidiasis, caused by the nematodes Strongyloides stercoralis and S. fuelleborni, is a neglected tropical disease affecting millions of people in tropical and subtropical areas. Non-human primates (NHPs) are typically natural hosts of S. fuelleborni but this parasite may spread to humans by zoonotic transmission. In this study, the presence of Strongyloides infections in NHPs in Khon Kaen Zoo, Thailand, was tested for using the agar plate culture technique. Three out of nine species of NHPs harbored S. fuelleborni (33.3%). The overall prevalence from three infected species was 45.5% (5 of 11 animals). DNA was amplified from larval stages of S. fuelleborni and a portion of the 18S ribosomal RNA gene was sequenced. These sequences were identical to previously published for S. fuelleborni. The parasitological and molecular data obtained in this study confirmed the presence of S. fuelleborni among captive NHPs (Bornean orangutans, chimpanzees, and red-shanked douc langurs) in Khon Kaen Zoo in Thailand. There is therefore high transmission potential of S. fuelleborni from NHP reservoir hosts to humans and the subsequent development of human strongyloidiasis.