Giardiasis is the most common enteric protozoan infection notifiable in New South Wales (NSW), Australia. Surveillance by NSW Health had shown a steady increase (prior to the COVID-19 pandemic) in the number of cases reported since 2012 and the reasons for this currently remain unknown. This study aimed to investigate the occurrence of Giardia intestinalis assemblages causing human infection in NSW. Individual faecal specimens were collected from participating hospitals and private laboratories, and the presence of Giardia and co-infections was confirmed by real-time multiplex-polymerase chain reaction (PCR). Samples were genotyped by sequence analysis of the triose phosphate isomerase (tpi) gene and the small subunit rDNA. Combined genotyping showed that most samples belong to assemblage B, and only a small percentage were infected with only assemblage A. Mixtures of assemblages A and B in individuals were relatively common. Co-infections were observed in ∼ half of the cases, with the most common co-infection being Blastocystis hominis and Dientamoeba fragilis. Although giardiasis was more prevalent in males, the assemblage distribution between the sexes appeared uniform. The age distribution was bimodal, with peaks in 0-15-year-olds and in adults in their 30s. The overall largest number of cases was collected from patients aged 30-49 years. Interestingly, females aged 5 years old and under had a greater risk of assemblage B infection than their male counterparts. No significant correlation was found between assemblage and clinical symptoms. This study provides new insights into the molecular diversity of giardiasis in NSW and helps inform enhanced surveillance and prevention strategies in Sydney.

Real-time PCR (qPCR) diagnostics developed for use in human clinical settings have been implemented to identify new animal hosts of the gastrointestinal protozoan Dientamoeba fragilis. The gut microbiome varies between species; unrecognised cross-reactivity could occur when applying these assays to new animal hosts. The use of qPCR diagnostics was assessed for the identification of new animal hosts of the gastrointestinal protozoan Dientamoeba fragilis. Forty-nine cattle, 84 dogs, 39 cats and 254 humans were screened for D. fragilis using two qPCR assays: EasyScreen (Genetic Signatures) and a laboratory-based assay commonly used in Europe. The reliability of the identifications made by these assays were assessed using melt curve analysis of qPCR products, conventional PCR targeting the SSU rDNA sequencing and NGS amplicon sequencing of qPCR product. PCR products from the D. fragilis identified in cattle had a 9 °C cooler melt curve than when detected in humans. This melt curve discrepancy, indicative of cross-reactivity with an unknown organism, was investigated further. DNA sequencing determined that Simplicimonas sp. was the genera responsible for this cross-reactivity in cattle specimens. Dientamoeba fragilis was not detected in either dogs or cats. There was a discrepancy in the number of positive samples detected using the two qPCR assays when applied to human samples. The EasyScreen assay detected 24 positive samples; the laboratory-based assay detected an additional 34 positive samples. Of the discrepant samples, 5 returned sequence data for D. fragilis, and 29 were unsupported (false) positive samples. Analysis of the melt curve after the qPCR reaction is a valuable technique to help differentiate samples containing D. fragilis compared to cross-reactions with non-target organisms. The identification of new animal hosts requires further evidence from either microscopy or DNA sequencing to confirm the presence of D. fragilis. Additionally, to reduce the risk of false-positive results due to non-specific amplification, we recommend reducing the number of PCR cycles to less than 40. Based on these results, we consider the ramifications of this identified cross-reactivity to the known host species distribution of D. fragilis.

Little is known about the life cycle and mode of transmission of Dientamoeba fragilis . Recently it was suggested that fecal–oral transmission of cysts may play a role in the transmission of D. fragilis . In order to establish an infection, D. fragilis is required to remain viable when exposed to the pH of the stomach. In this study, we investigated the ability of cultured trophozoites to withstand the extremes of pH. We provide evidence that trophozoites of D. fragilis are vulnerable to highly acidic conditions. We also investigated further the ultrastructure of D. fragilis cysts obtained from mice and rats by transmission electron microscopy. These studies of cysts showed a clear cyst wall surrounding an encysted parasite. The cyst wall was double layered with an outer fibrillar layer and an inner layer enclosing the parasite. Hydrogenosomes, endoplasmic reticulum and nuclei were present in the cysts. Pelta-axostyle structures, costa and axonemes were identifiable and internal flagellar axonemes were present. This study therefore provides additional novel details and knowledge of the ultrastructure of the cyst stage of D. fragilis .

An abstract is not available for this content so a preview has been provided. As you have access to this content, a full PDF is available via the 'Save PDF' action button.

Bibliometric and bibliographic analyses are popular tools for investigating publication metrics and thematic transitions in an expanding codex of biomedical literature. Bibliometric techniques have been employed in parasitology and vaccinology, with only a few malaria-specific literature analyses being reported specifically on parasite vaccines. The pursuit of parasite prophylactics is an important, global endeavour both medically and economically. As such, a comprehensive understanding of the research topics would be a valuable tool in assessing the current status and future directions of parasite vaccine development. Consequently, this study investigated parasite vaccinology from 1990 to 2019 by analysing literature exported from the Web of Science and Dimensions databases using two, commonly used, bibliometric programs: SciMAT and VOSviewer. The results of this study show the common, emerging, and transient themes within the discipline, and where the future lies as vaccine development moves further into the age of omics and informatics.

Reverse vaccinology (RV) was described at its inception in 2000 as an in silico process that starts from the genomic sequence of the pathogen and ends with a list of potential protein and/or peptide candidates to be experimentally validated for vaccine development. Twenty-two years later, this process has evolved from a few steps entailing a handful of bioinformatics tools to a multitude of steps with a plethora of tools. Other in silico related processes with overlapping workflow steps have also emerged with terms such as subtractive proteomics, computational vaccinology, and immunoinformatics. From the perspective of a new RV practitioner, determining the appropriate workflow steps and bioinformatics tools can be a time consuming and overwhelming task, given the number of choices. This review presents the current understanding of RV and its usage in the research community as determined by a comprehensive survey of scientific papers published in the last seven years. We believe the current mainstream workflow steps and tools presented here will be a valuable guideline for all researchers wanting to apply an up-to-date in silico vaccine discovery process.

Vaccine discovery against eukaryotic parasites is not trivial as highlighted by the limited number of known vaccines compared to the number of protozoal diseases that need one. Only three of 17 priority diseases have commercial vaccines. Live and attenuated vaccines have proved to be more effective than subunit vaccines but adversely pose more unacceptable risks. One promising approach for subunit vaccines is in silico vaccine discovery, which predicts protein vaccine candidates given thousands of target organism protein sequences. This approach, nonetheless, is an overarching concept with no standardised guidebook on implementation. No known subunit vaccines against protozoan parasites exist as a result of this approach, and consequently none to emulate. The study goal was to combine current in silico discovery knowledge specific to protozoan parasites and develop a workflow representing a state-of-the-art approach. This approach reflectively integrates a parasite’s biology, a host's immune system defences, and importantly, bioinformatics programs needed to predict vaccine candidates. To demonstrate the workflow effectiveness, every Toxoplasma gondii protein was ranked in its capacity to provide long-term protective immunity. Although testing in animal models is required to validate these predictions, most of the top ranked candidates are supported by publications reinforcing our confidence in the approach.

This study investigated the emergence and use of Twitter, as of July 2023 being rebranded as X, as the main forum for social media communication in parasitology. A dataset of tweets was constructed using a keyword search of Twitter with the search terms ‘malaria’, ‘Plasmodium’, ‘Leishmania’, ‘Trypanosoma’, ‘Toxoplasma’ and ‘Schistosoma’ for the period from 2011 to 2020. Exploratory data analyses of tweet content were conducted, including language, usernames and hashtags. To identify parasitology topics of discussion, keywords and phrases were extracted using KeyBert and biterm topic modelling. The sentiment of tweets was analysed using VADER. The results show that the number of tweets including the keywords increased from 2011 (for malaria) and 2013 (for the others) to 2020, with the highest number of tweets being recorded in 2020. The maximum number of yearly tweets for Plasmodium, Leishmania, Toxoplasma, Trypanosoma and Schistosoma was recorded in 2020 (2804, 2161, 1570, 680 and 360 tweets, respectively). English was the most commonly used language for tweeting, although the percentage varied across the searches. In tweets mentioning Leishmania, only ∼37% were in English, with Spanish being more common. Across all the searches, Portuguese was another common language found. Popular tweets on Toxoplasma contained keywords relating to mental health including depression, anxiety and schizophrenia. The Trypanosoma tweets referenced drugs (benznidazole, nifurtimox) and vectors (bugs, triatomines, tsetse), while the Schistosoma tweets referenced areas of biology including pathology, eggs and snails. A wide variety of individuals and organisations were shown to be associated with Twitter activity. Many journals in the parasitology arena regularly tweet about publications from their journal, and professional societies promote activity and events that are important to them. These represent examples of trusted sources of information, often by experts in their fields. Social media activity of influencers, however, who have large numbers of followers, might have little or no training in science. The existence of such tweeters does raise cause for concern to parasitology, as one may start to question the quality of information being disseminated.

Neglected tropical diseases affect those in poorer nations disproportionately across the globe. One example of these, leishmaniasis, is a debilitating and potentially fatal parasitic infection. Molecular detection of this disease can provide accurate and fast diagnosis, and with near point-of-care technologies, detection can be provided in many health-care settings. Traditionally, the perceived limitations to such detection methods have hindered their provision to resource-limited nations, but new technologies and techniques are helping to overcome these perceptions. The current pandemic offers an opportunity to maintain and develop further advances, ensuring molecular diagnostics are accessible to all.

The change is not trivial for our current financial plan, primarily based upon institutional subscriptions, is transformed to an article processing charges (APCs) one.In order to accommodate peer review and production lead times, all papers submitted after 3rd October 2022, if accepted, will be published as OA.By transitioning to OA, Parasitology seeks to improve the global accessibility of its articles, with a more equitable international readership, and responds to the behests of research funders requiring authors to publish in OA journals.All submissions to Parasitology will continue to be fully peer-reviewed to maintain our high standards.With respect to our on-going attention on research integrity and quality assurance processes, we have adopted the use of iThenicate (see https://www.ithenticate.com/)with Crossref providing a database of scholarly papers.Using these e-tools, we can better detect any evidence of plagiarism within forthcoming manuscripts once assigned for peer review.We stress that evidence of plagiarism within our journal is very rare, but these steps are taken from preventive rather than retrospective concerns.Many of you are likely familiar with its sister technology Turnitin (see https://www.turnitin.com/)that is regularly used in academic institutions to search for plagiarism in student essays and assessments.Our use of iThenicate aligns us with other high-profile publishers such as Springer Nature, Wiley and Elsevier, and also helps us to better assess the novelty of work submitted.

Leishmania infections span a range of clinical syndromes and impact humans from many geographic foci, but primarily the world's poorest regions. Transmitted by the bite of a female sand fly, Leishmania infections are increasing with human movement (due to international travel and war) as well as with shifts in vector habitat (due to climate change). Accurate diagnosis of the 20 or so species of Leishmania that infect humans can lead to the successful treatment of infections and, importantly, their prevention through modelling and intervention programs. A multitude of laboratory techniques for the detection of Leishmania have been developed over the past few decades, and although many have drawbacks, several of them show promise, particularly molecular methods like polymerase chain reaction. This review provides an overview of the methods available to diagnostic laboratories, from traditional techniques to the now-preferred molecular techniques, with an emphasis on polymerase chain reaction-based detection and typing methods.

Given the importance of angiostrongyliasis as an emerging infectious disease of humans, companion animals, and wildlife, the current study focused on the transmission dynamics of first- and third-stage larvae of the parasitic nematode, Angiostrongylus cantonensis . The migration of infective larvae and their subsequent distribution within the Lymnaeidae snail, Bullastra lessoni , were investigated over time using microscopic examination of histological sections and fresh tissue. Snails were divided into four anatomical regions: (i) anterior and (ii) posterior cephalopedal masses, (iii) mantle skirt and (iv) visceral mass. The viability of free-swimming third-stage larvae, after their release from snail tissues, was evaluated in vitro by propidium iodide staining and infectivity by in vivo infection of Wistar rats. Snails were sequentially dissected over time to assess the number and anatomical distribution of larvae within each snail and hence infer their migration pathway. Herein, ongoing larval migratory activity was detected over 28 days post-infection. A comparison of infection rates and the larval distribution within the four designated snail regions demonstrated a significant relationship between anatomical region and density of infective larvae, with larvae mostly distributed in the anterior cephalopedal mass (43.6 ± 10.8%) and the mantle skirt (33.0 ± 8.8%). Propidium iodide staining showed that free-swimming third-stage larvae retained viability for between 4 and 8 weeks when stored under laboratory conditions. In contrast to viability, larval infectivity in rats remained for up to 2 weeks only. Knowledge gained from the current work could provide information on the development of new approaches to controlling the transmission of this parasite.

The World Health Organisation reported in 2020 that six of the top 10 sources of death in low-income countries are parasites. Parasites are microorganisms in a relationship with a larger organism, the host. They acquire all benefits at the host’s expense. A disease develops if the parasitic infection disrupts normal functioning of the host. This disruption can range from mild to severe, including death. Humans and livestock continue to be challenged by established and emerging infectious disease threats. Vaccination is the most efficient tool for preventing current and future threats. Immunogenic proteins sourced from the disease-causing parasite are worthwhile vaccine components (subunits) due to reliable safety and manufacturing capacity. Publications with ‘subunit vaccine’ in their title have accumulated to thousands over the last three decades. However, there are possibly thousands more reporting immunogenicity results without mentioning ‘subunit’ and/or ‘vaccine’. The exact number is unclear given the non-standardised keywords in publications. The study aim is to identify parasite proteins that induce a protective response in an animal model as reported in the scientific literature within the last 30 years using machine learning and natural language processing. Source code to fulfil this aim and the vaccine candidate list obtained is made available.

Deletion of histidine-rich protein genes pfhrp2/3 in Plasmodium falciparum causes infections to go undetected by HRP2-based malaria rapid diagnostic tests. We analyzed P. falciparum malaria cases imported to Australia (n = 210, collected 2010-2018) for their pfhrp2/3 status. We detected gene deletions in patients from 12 of 25 countries. We found >10% pfhrp2-deletion levels in those from Nigeria (13.3%, n = 30), Sudan (11.2%, n = 39), and South Sudan (17.7%, n = 17) and low levels of pfhrp3 deletion from Sudan (3.6%) and South Sudan (5.9%). No parasites with pfhrp2/3 double deletions were detected. Microsatellite typing of parasites from Nigeria, Sudan, and South Sudan revealed low relatedness among gene-deleted parasites, indicating independent emergences. The gene deletion proportions signify a risk of false-negative HRP2-RDT results. This study's findings warrant surveillance to determine whether the prevalence of gene-deleted parasites justifies switching malaria rapid diagnostic tests in Nigeria, Sudan, and South Sudan.

Babesia infection of red blood cells can cause a severe disease called babesiosis in susceptible hosts. Bovine babesiosis causes global economic loss to the beef and dairy cattle industries, and canine babesiosis is considered a clinically significant disease. Potential therapeutic targets against bovine and canine babesiosis include members of the exportome, i.e., those proteins exported from the parasite into the host red blood cell. We developed three machine learning-derived methods (two novel and one adapted) to predict for every known Babesia bovis, Babesia bigemina, and Babesia canis protein the probability of being an exportome member. Two well-studied apicomplexan-related species, Plasmodium falciparum and Toxoplasma gondii, with extensive experimental evidence on their exportome or excreted/secreted proteins were used as important benchmarks for the three methods. Based on 10-fold cross validation and multiple train–validation–test splits of training data, we expect that over 90% of the predicted probabilities accurately provide a secretory or non-secretory indicator. Only laboratory testing can verify that predicted high exportome membership probabilities are creditable exportome indicators. However, the presented methods at least provide those proteins most worthy of laboratory validation and will ultimately save time and money.

Giardia intestinalis continues to be one of the most encountered parasitic diseases around the world. Although more frequently detected in developing countries, Giardia infections nonetheless pose significant public health problems in developed countries as well. Molecular characterisation of Giardia isolates from humans and animals reveals that there are two genetically different assemblages (known as assemblage A and B) that cause human infections. However, the current molecular assays used to genotype G. intestinalis isolates are quite controversial. This is in part due to a complex phenomenon where assemblages are incorrectly typed and underreported depending on which targeted locus is sequenced. In this review, we outline current knowledge based on molecular epidemiological studies and raise questions as to the reliability of current genotyping assays and a lack of a globally accepted method. Additionally, we discuss the clinical symptoms caused by G. intestinalis infection and how these symptoms vary depending on the assemblage infecting an individual. We also introduce the host-parasite factors that play a role in the subsequent clinical presentation of an infected person, and explore which assemblages are most seen globally.

Bioinformatics programs have been developed that exploit informative signals encoded within protein sequences to predict protein characteristics. Unfortunately, there is no program as yet that can predict whether a protein will induce a protective immune response to a pathogen. Nonetheless, predicting those pathogen proteins most likely from those least likely to induce an immune response is feasible when collectively using predicted protein characteristics. Vacceed is a computational pipeline that manages different standalone bioinformatics programs to predict various protein characteristics, which offer supporting evidence on whether a protein is secreted or membrane -associated. A set of machine learning algorithms predicts the most likely pathogen proteins to induce an immune response given the supporting evidence. This chapter provides step by step descriptions of how to configure and operate Vacceed for a eukaryotic pathogen of the user’s choice.