Morphological and molecular characterization of Eimeria haematopusi n. sp. (Apicomplexa: Eimeriidae) in an Australian Pied Oystercatcher (Haematopus longirostris) (Aves: Charadriiformes).

A novel Eimeria Schneider, 1875 species is described from an Australian pied oystercatcher Haematopus longirostris Vieillot, in Western Australia. The pied oystercatcher was admitted to the Kanyana Wildlife Rehabilitation Centre (KWRC), Perth, Western Australia in a poor body condition, abrasion to its right hock and signs of partial delamination to its lower beak. Investigation into potential medical causes resulted in a faecal sample being collected and screened for gastrointestinal parasites. Unsporulated coccidian oocysts were initially observed in the faeces and identified as Eimeria upon sporulation. The sporulated oocysts (n = 20) are ellipsoidal, 20-21 × 12-13 µm in shape and have thick bi-layered walls which are c.2/3 of the total thickness. Micropyle is present, robust and protruding, and occasionally has a rounded polar body attached to the micropyle. Within the oocyst, a residuum, in addition, two to five polar granules are present. There are four ellipsoidal sporocysts 9-11 × 5-6 µm with flattened to half-moon shaped Stieda bodies. Sub-Stieda body and para-Stieda body are absent. The sporocysts contain sporocyst residuums composed of a few spherules scattered among the sporozoites. Within the sporozoites, anterior and posterior refractile bodies are present, but the nucleus is indiscernible. To further characterise the novel Eimeria species from H. longirostris, molecular analysis was conducted at the 18S ribosomal RNA locus, using PCR amplification and cloning. Two cloned sequences from the novel Eimeria were compared with those from other Eimeria spp. with the highest genetic similarity of 97.6% and 97.2% from Clone 1 and 2, respectively with Eimeria reichenowi (AB544308) from a hooded crane (Grus monacha Temminck) in Japan. Both sequences grouped in a clade with the Eimeria spp. isolated from wetland birds, which include Eimeria paludosa (KJ767187) from a dusky moorhen (Gallinula tenebrosa Gould) in Western Australia, Eimeria reichenowi (AB544308) and Eimeria gruis (AB544336) both from hooded cranes. Based on the morphological and molecular data, this Eimeria sp. is a new species of coccidian parasite and is named Eimeria haematopusi n. sp. after its host H. longirostris.

Next-generation sequencing amplicon analysis of the genetic diversity of Eimeria populations in livestock and wildlife samples from Australia.

Eimeria is an important coccidian enteric parasite that infects a wide range of hosts and can cause substantial economic losses in the poultry and livestock industries. It is common for multiple Eimeria species to infect individual hosts, and this can make species identification difficult due to morphological similarities between species and mixed chromatograms when using Sanger sequencing. Relatively few studies have applied next-generation amplicon sequencing (NGS) to determining the genetic diversity of Eimeria species in different hosts. The present study screened 408 faecal samples from a range of hosts including livestock and wildlife using a previously developed quantitative polymerase chain reaction (qPCR) at the 18S locus and conducted amplicon NGS on the positives using a ~ 455-bp fragment of the 18S locus. A total of 41 positives (10.1%) were identified by qPCR from various hosts and NGS was successful for 38 of these positives. Fifteen Eimeria species and three genotypes were detected by NGS: E. ferrisi, E. kanyana, E. potoroi, E. quokka, E. setonicis, E. trichosuri, E. reichenowi, E. angustus, E. ahsata, E. auburnensis, E. bovis, E. brasiliensis, E. christenseni, E. crandallis, E. ovinoidalis, Eimeria sp. (JF419345), Eimeria sp. (JF419349) and Eimeria sp. (JF419351). Mixed infections were detected in 55.3% (21/38) of positive samples. The most striking finding was the identification of the same species in different hosts. This could be due to contamination and/or mechanical transmission or may provide support for previous studies suggesting that Eimeria species can infect not just closely related hosts but different genera and further research is required. This is also the first study to audit Eimeria populations in livestock (sheep and cattle) by NGS and could be applied in the future to determine the extent of pathogenic species and outcomes of Eimeria control strategies.

Cryptosporidium abrahamseni n. sp. (Apicomplexa: Cryptosporidiiae) from red-eye tetra (Moenkhausia sanctaefilomenae).

The morphological, biological, and molecular characterisation of Cryptosporidium piscine genotype 7 from red-eye tetras (Moenkhausia sanctaefilomenae) are described, and the species name Cryptosporidium abrahamseni n. sp. is proposed. Histological analysis of intestinal tissue identified large numbers of Cryptosporidium organisms along the epithelial lining of the intestine. Sequence and phylogenetic analysis at 18S rRNA (18S) and actin loci conducted on intestinal scrapings revealed that C. abrahamseni n. sp. was genetically distinct from other Cryptosporidium species. At the 18S locus, it was most closely related to C. huwi (3.2% genetic distance) and exhibited genetic distances ranging from 5.9 to 6.5% (C. molnari) to 14.9% (C. scolpthalmi) from all other Cryptosporidium species. At the actin locus, the genetic distances were larger and C. abrahamseni n. sp. exhibited 10.3% genetic distance from C. huwi, and 17.6% (C. molnari) to 28% (C. canis) genetic distance from other Cryptosporidium spp. Phylogenetic analysis of concatenated 18S and actin sequences confirmed that C. abrahamseni n. sp. shares the closest genetic relationship with C. huwi (6.7% genetic distance), while the genetic distance between C. abrahamseni n. sp. and other Cryptosporidium spp. ranged from 12.1% (C. molnari) to 20.4% (C. canis). Based on genetic and histological data, C. abrahamseni n. sp. is validated as a separate species.

Wastewater-based epidemiology-surveillance and early detection of waterborne pathogens with a focus on SARS-CoV-2, Cryptosporidium and Giardia.

Waterborne diseases are a major global problem, resulting in high morbidity and mortality, and massive economic costs. The ability to rapidly and reliably detect and monitor the spread of waterborne diseases is vital for early intervention and preventing more widespread disease outbreaks. Pathogens are, however, difficult to detect in water and are not practicably detectable at acceptable concentrations that need to be achieved in treated drinking water (which are of the order one per million litre). Furthermore, current clinical-based surveillance methods have many limitations such as the invasive nature of the testing and the challenges in testing large numbers of people. Wastewater-based epidemiology (WBE), which is based on the analysis of wastewater to monitor the emergence and spread of infectious disease at a population level, has received renewed attention in light of the current coronavirus disease 2019 (COVID-19) pandemic. The present review will focus on the application of WBE for the detection and surveillance of pathogens with a focus on severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and the waterborne protozoan parasites Cryptosporidium and Giardia. The review highlights the benefits and challenges of WBE and the future of this tool for community-wide infectious disease surveillance.

Cryptosporidium bollandi n. sp. (Apicomplexa: Cryptosporidiiae) from angelfish (Pterophyllum scalare) and Oscar fish (Astronotus ocellatus).

The species name Cryptosporidium bollandi n. sp. is proposed for Cryptosporidium piscine genotype 2 based on morphological, biological and molecular characterisation. Phylogenetic analyses of 18S rRNA (18S) sequences revealed that C. bollandi n. sp. was most closely related to piscine genotype 4 (5.1% genetic distance) and exhibited genetic distances of 10.0%, 12.2% and 25.2% from Cryptosporidium molnari, Cryptosporidium huwi and Cryptosporidium scophthtalmi, respectively. At the actin locus, C. bollandi n. sp. was again most closely related to piscine genotype 4 (6.8% genetic distance) and exhibited 15.5% (C. molnari), 18.4% (C. huwi), 22.9% (C. scophthalmi) and up to 27.5% genetic distance from other Cryptosporidium spp. (Cryptosporidium felis). Phylogenetic analysis of concatenated 18S and actin sequences showed that C. bollandi n. sp. exhibited 12.9% (C. molnari) to 21.1% (C. canis) genetic distance from all other Cryptosporidium spp. Genetic data as well as previous histological analysis clearly supports the validity of C. bollandi n. sp. as a separate species.

Further characterisation of Haemocystidium chelodinae-like Haemoproteidae isolated from the Bellinger River snapping turtle (Myuchelys georgesi).

The Bellinger River snapping turtle (Myuchelys georgesi) is endemic to Australia and is confined to a highly restricted distribution in the Bellinger River in New South Wales. Routine veterinary health examinations of 17 healthy turtles were undertaken, along with the collection and analysis of blood samples, during conservation efforts to save the species following a catastrophic population decline. Microscopy analysis of blood films detected Haemoproteidae parasites that morphologically resembled Haemocystidium chelodinae inside turtle erythrocytes. Of the 17 turtles examined, 16 were positive for infection with H. chelodinae by both light microscopy and PCR. DNA sequencing of a partial fragment of the mitochondrial cytochrome b (cytb) gene and phylogenetic analysis identified two different H. chelodinae-like genotypes. The phylogenetic relationship of H. chelodinae-like to other Haemoproteidae species based on cytb sequences grouped H. chelodinae-like into the reptile clade, but revealed the Haemocystidium genus to be paraphyletic as the clade also contained Haemoproteus, thus supporting a re-naming of Haemoproteus species from reptiles to Haemocystidium species. This study reports for the first time the genetic characterisation of H. chelodinae-like organisms isolated from a new Testudine host species, the Bellinger River snapping turtle. As evidence grows, further research will be necessary to understand the mode of transmission and to investigate whether these parasites are pathogenic to their hosts.

Molecular typing of Giardia duodenalis in humans in Queensland - first report of Assemblage E.

Little is known about the genetic diversity of the protozoan parasite, Giardia duodenalis, infecting humans in Queensland, Australia. The present study typed 88 microscopically Giardia-positive isolates using assemblage-specific primers at the triose phosphate isomerase (tpi) gene and sequenced a subset of isolates at the glutamate dehydrogenase (gdh) gene (n = 30) and tpi locus (n = 27). Using the tpi-assemblage specific primers, G. duodenalis assemblage A and assemblage B were detected in 50% (44/88) and 38·6% (34/88) of samples, respectively. Mixed infections with assemblages A and B were identified in 4·5% (4/88) and assemblage E was identified in 6·8% (6/88) of samples. Sequence analysis at the gdh and tpi loci also confirmed the presence of assemblage E in these isolates. Cyst numbers per gram of feces (g-1) were determined using quantitative polymerase chain reaction and of the isolates that were typed as assemblage E, cyst numbers ranged 13·8-68·3 × 106 cysts g-1. This is the first report of assemblage E in humans in Australia, indicating that in certain settings, this assemblage may be zoonotic.

First report of Cryptosporidium species in farmed and wild buffalo from the Northern Territory, Australia.

A molecular epidemiological survey of Cryptosporidium from water buffalo (Bubalus bubalis) in the Northern Territory in Australia was conducted. Fecal samples were collected from adult farmed (n = 50) and wild buffalo (n = 50) and screened using an 18S quantitative PCR (qPCR). Positives were typed by sequence analysis of 18S nested PCR products. The qPCR prevalence of Cryptosporidium species in farmed and wild buffalo was 30 and 12 %, respectively. Sequence analysis identified two species: C. parvum and C. bovis, with C. parvum accounting for ~80 % of positives typed from the farmed buffalo fecal samples compared to 50 % for wild buffalo. Subtyping at the 60 kDa glycoprotein (gp60) locus identified C. parvum subtypes IIdA19G1 (n = 4) and IIdA15G1 (n = 1) in the farmed buffalo and IIaA18G3R1 (n = 2) in the wild buffalo. The presence of C. parvum, which commonly infects humans, suggests that water buffaloes may contribute to contamination of rivers and waterways with human infectious Cryptosporidium oocysts, and further research on the epidemiology of Cryptosporidium in buffalo populations in Australia is required.

Identification and genetic characterization of a novel species of Choleoeimeria Schneider, 1875 from a captive-bred bilby (Thylacomyidae; Macrotis lagotis) (Reid, 1837) in Western Australia.

A novel Eimeria sp. from a captive-bred bilby (Macrotis lagotis Reid, 1837) has been identified in Western Australia. The bilby was bred at the Kanyana Wildlife Rehabilitation Centre, Perth, as part of the National Bilby Recovery Plan. Oocysts (n = 31) irregular blunt ellipsoidal, 17-18 × 11-12 (17.2 × 11.3); length/width (L/W) ratio 1.4-1.5 (1.5). Wall bi-layered, 0.8-1.0 (0.9) thick, outer layer smooth, c.2/3 of total thickness. Micropyle barely discernible. Oocyst residuum is absent, but 2-3 small polar granules are present. Sporocysts (n = 31) ovoidal, 7-8 × 5-6 (7.8 × 5.7); L/W ratio 1.3-1.4 (1.4). Stieda, sub-Stieda and para-Stieda bodies absent or indiscernible; sporocyst residuum present, usually as an irregular body consisting of numerous granules that appear to be membrane-bound or sometimes diffuse among sporozoites. Sporozoites vermiform with a robust refractile body. Further molecular characterization was conducted on the sporulated oocysts. At the 18S locus, it sat in a large clade of the phylogenetic tree with two isolates of Eimeria angustus from quendas (Isoodon obesulus Shaw, 1797) and the Choleoeimeria spp. It shared the highest identity with E. angustus (KU248093) at 98.84%; at the COI gene locus, it was unique and most closely related to Choleoeimeria taggarti, which is hosted by another species of marsupial, the yellow-footed antechinus (Antechinus flavipes flavipes), with 90.58% genetic similarity. Based on morphological and molecular data, this isolate is a new species and named as Choleoeimeria yangi n. sp.

Efficacy and safety of alpha-pinene capsule in the management of functional dyspepsia and eradication of helicobacter pylori: a randomized clinical trial.

Aim: Our objective was to assess the efficacy and safety of adding alpha-pinene (a herbal terpenoid) to quadruple therapy compared to a placebo in improving symptoms and Helicobacter pylori (H. pylori) eradication rates in Functional dyspepsia (FD) patients. Background: FD is a prevalent upper gastrointestinal condition, and no definitive pharmacological treatment is available for its management. Methods: We conducted a randomized, double-blinded, placebo-controlled trial on FD patients diagnosed with H. pylori infection. We collected baseline demographic data and assessed FD symptoms in the participants. Patients were randomly allocated to receive either standard quadruple therapy with α-pinene capsules (0.25 mg/day) or quadruple therapy with a placebo for two weeks. We employed a validated questionnaire, the Short Form Leeds Dyspepsia Questionnaire (SF-LDQ), to evaluate FD symptoms. The eradication rate of H. pylori was compared between the two groups one month after completing the treatment regimens. Any reported adverse drug reactions (ADRs) were documented throughout the trial. Results: Over four months, a total of 66 patients completed the trial. Notably, there were no significant differences in baseline SF-LDQ scores between the two groups (p=0.83); however, a significant divergence emerged at the trial's conclusion (p=0.03). The H. pylori eradication rates did not show notable differences between the two treatment arms (p=0.43). Importantly, there were no dropouts from the trial due to ADRs. Among reported ADRs, participants experienced abdominal pain, headache, diarrhea, and a metallic taste, with no significant variance in incidence rates observed between the two groups (p=0.62). Conclusion: These findings suggest that α-pinene could be an effective and safe agent for reducing FD symptoms.

Medicinal plants as a source of antiparasitics: an overview of experimental studies.

Despite advances in modern human and veterinary medicine, gastrointestinal (GI) parasitic infections remain a significant health issue worldwide, mainly in developing countries. Increasing evidence of the multi-drug resistance of these parasites and the side effects of currently available synthetic drugs have led to increased research on alternative medicines to treat parasitic infections. The exploration of potential botanical antiparasitics, which are inexpensive and abundant, may be a promising alternative in this context. This study summarizes the in vitro/in vivo antiparasitic efficacy of different medicinal plants and their components against GI parasites. Published literature from 1990-2020 was retrieved from Google Scholar, Web of Science, PubMed and Scopus. A total of 68 plant species belonging to 32 families have been evaluated as antiparasitic agents against GI parasites worldwide. The majority of studies (70%) were conducted in vitro. Most plants were from the Fabaceae family (53%, n = 18). Methanol (37%, n = 35) was the most used solvent. Leaf (22%, n = 16) was the most used plant part, followed by seed and rhizome (each 12%, n = 9). These studies suggest that herbal medicines hold a great scope for new drug discoveries against parasitic diseases and that the derivatives of these plants are useful structures for drug synthesis and bioactivity optimization.

Mitigation approaches and techniques for combustion power plants flue gas emissions: A comprehensive review.

Population growth and urbanization are driving energy demand. Despite the development of renewable energy technologies, most of this demand is still met by fossil fuels. Flue gases are the main air pollutants from combustion power plants. These pollutants include particulate matter (PM), sulfur oxides (SOx), nitrogen oxides (NOx), and carbon oxides (COx). The release of these pollutants has adverse effects on human health and the environment, including serious damage to the human respiratory system, acid rain, climate change, and global warming. In this review, a wide range of conventional and new technologies that have the potential to be used in the combustion power plant sector to manage and reduce flue gas pollutants have been examined. Nowadays, conventional approaches to emissions control and management, which focus primarily on post-combustion techniques, face several challenges despite their widespread use and commendable effectiveness. Therefore, studies that have proposed alternative approaches to achieve improved and more efficient methods are reviewed. The results show that new advances such as novel PM collectors, attaining an efficiency of nearly 100 % for submicron particles, microwave systems, boasting an efficiency of nearly 90 % for NO and over 95 % for SO2, electrochemical systems achieving above 90 % efficiency for NOx reduction, non-thermal plasma processes demonstrating an efficiency close to 90 % for NOx, microalgae-based methods with efficiency ranging from 80 % to 99 % for CO2, and wet scrubbing, exhibit considerable potential in addressing the shortcomings of conventional systems. Furthermore, the integration of hybrid methods, particularly in regions prioritizing environmental concerns over economic considerations, holds promise for enhanced control and removal of flue gas pollutants with superior efficiency.

Economic evaluations of whole-genome sequencing for pathogen identification in public health surveillance and health-care-associated infections: a systematic review.

Whole-genome sequencing (WGS) has resulted in improvements to pathogen characterisation for the rapid investigation and management of disease outbreaks and surveillance. We conducted a systematic review to synthesise the economic evidence of WGS implementation for pathogen identification and surveillance. Of the 2285 unique publications identified through online database searches, 19 studies met the inclusion criteria. The economic evidence to support the broader application of WGS as a front-line pathogen characterisation and surveillance tool is insufficient and of low quality. WGS has been evaluated in various clinical settings, but these evaluations are predominantly investigations of a single pathogen. There are also considerable variations in the evaluation approach. Economic evaluations of costs, effectiveness, and cost-effectiveness are needed to support the implementation of WGS in public health settings.

In Vitro Susceptibility of Cryptosporidium parvum to Plant Antiparasitic Compounds.

Cryptosporidium parvum is a significant cause of watery diarrhoea in humans and other animals worldwide. Although hundreds of novel drugs have been evaluated, no effective specific chemotherapeutic intervention for C. parvum has been reported. There has been much recent interest in evaluating plant-derived products in the fight against gastrointestinal parasites, including C. parvum. This study aimed to identify extracts from 13 different plant species that provide evidence for inhibiting the growth of C. parvum in vitro. Efficacy against C. parvum was detected and quantified using quantitative PCR and immunofluorescence assays. All plant extracts tested against C. parvum showed varying inhibition activities in vitro, and none of them produced a cytotoxic effect on HCT-8 cells at concentrations up to 500 µg/mL. Four plant species with the strongest evidence of activity against C. parvum were Curcuma longa, Piper nigrum, Embelia ribes, and Nigella sativa, all with dose-dependent efficacy. To the authors' knowledge, this is the first time that these plant extracts have proven to be experimentally efficacious against C. parvum. These results support further exploration of these plants and their compounds as possible treatments for Cryptosporidium infections.

Eimeria spp. and Tyzzeria perniciosa Allen, 1936 (Apicomplexa: Eimeriidae) from a Pacific black duck, Anas superciliosa Gmelin (Aves: Anseriformes), in western Australia.

Four species of the Eimeriidae, Eimeria anatis Scholtyseck, 1955, Eimeria aythyae Farr, 1965, Eimeria krylovi Svanbaev & Rakhmatullina, 1967 and Tyzzeria perniciosa Allen, 1936, were morphologically identified from oöcysts recovered from a Pacific black duck, Anas superciliosa Gmelin. Additionally, genotypic characterization of E. anatis is provided via sequencing of the mitochondrial cytochrome c oxidase subunit 1 (cox1) and the small subunit ribosomal RNA (18S) genes. The four species are redescribed, providing additional morphological details. The validity of genera and coccidian species parasitizing birds of the order Anseriformes such as Wenyonella Hoare, 1933 and some Tyzzeria spp. are discussed. Molecular phylogenetic analyses for the cox1 and 18S rRNA genes resulted in monophylies of Eimeria spp. from Anseriformes which included the sequences obtained from E. anatis oöcysts.

Knowledge, Attitude and Practices Towards Cryptosporidium Among Public Swimming Pool Patrons and Staff in Western Australia.

PURPOSE: There is a dearth of research conducted on the Knowledge, Attitude and Practices (KAP) of swimming pool patrons and staff to determine their understanding of the importance of Cryptosporidium and its transmission in swimming pools. METHODS: We conducted a KAP survey of public swimming pool patrons (n = 380) and staff (n = 40) attending five public swimming pools in Western Australia (WA). RESULTS: Knowledge, attitudes and practices (KAP) of Cryptosporidium varied between patrons and staff but were generally limited. Only 26.1% and 25.0% of patrons and staff had heard of Cryptosporidium, while 17.4% and 10.0% knew that it causes diarrhoea, respectively. Thirty-one percent of patrons were aware of their pool policy concerning gastroenteritis and Cryptosporidium, compared to 62.5% of staff. Less than 50% of patrons demonstrated awareness of how features within the pool environment were relevant to the control of Cryptosporidium. Only about a third of patrons (35%) and staff (37.5%) were aware that showering before swimming reduced the risk of gastroenteritis. CONCLUSION: Raising awareness about hygiene-related practices through the delivery of targeted health education messages to the general public is essential to reduce the burden of Cryptosporidium infections in aquatic environments.

An Update on Zoonotic Cryptosporidium Species and Genotypes in Humans.

The enteric parasite, Cryptosporidium is a major cause of diarrhoeal illness in humans and animals worldwide. No effective therapeutics or vaccines are available and therefore control is dependent on understanding transmission dynamics. The development of molecular detection and typing tools has resulted in the identification of a large number of cryptic species and genotypes and facilitated our understanding of their potential for zoonotic transmission. Of the 44 recognised Cryptosporidium species and >120 genotypes, 19 species, and four genotypes have been reported in humans with C. hominis, C. parvum, C. meleagridis, C. canis and C. felis being the most prevalent. The development of typing tools that are still lacking some zoonotic species and genotypes and more extensive molecular epidemiological studies in countries where the potential for transmission is highest are required to further our understanding of this important zoonotic pathogen. Similarly, whole-genome sequencing (WGS) and amplicon next-generation sequencing (NGS) are important for more accurately tracking transmission and understanding the mechanisms behind host specificity.

Molecular characterization of Entamoeba, Blastocystis and Cryptosporidium species in stool samples collected from Jordanian patients suffering from gastroenteritis.

Little is known about the prevalence of intestinal protozoa in patients suffering from diarrhea in Jordan. The present study aimed to detect and speciate Entamoeba, Blastocystis, and Cryptosporidium species in a total of 159 human patients with diarrhea from November 2014 to October 2016. The overall prevalence for the three parasites was 19.5% (31/159). Entamoeba spp. (Entamoeba. dispar and/or Entamoeba histolytica), Blastocystis hominis, and Cryptosporidium parvum subtype IIaA15G2R1 were detected in 12.6%, 6%, and 0.6 of samples, respectively. This is the first molecular study in Jordan to confirm the diagnosis of Entamoeba species and to discriminate between E. histolytica and E. dispar.

Morphological and molecular characterization of a new species of Isospora Schneider, 1881 (Apicomplexa: Eimeriidae) from the western wattlebird Anthochaera lunulata Gould in Western Australia.

A new coccidian species, Isospora lunulatae n. sp., from the western wattlebird Anthochaera lunulata Gould in Western Australia is described and characterised molecularly. Microscopic analysis of a faecal sample identified subspheroidal oöcysts measuring 27-34 × 26-31 (30.6 × 29.4) µm (n = 20), with a length/width (L/W) ratio of 1.0-1.1 (1.0). Oöcysts have a bi-layered wall, 0.9-1.2 (1.0) µm thick; the outer layer is smooth, representing c.2/3 of total thickness. Micropyle and oöcyst residuum are both absent, but a polar granule is present. Sporocysts are ovoidal, 17-19 × 10-12 (18.3 × 10.7) µm, with a L/W ratio of 1.6-1.8 (1.7) and occupying about 21% of the area (each one) within the oöcyst. Stieda body is flattened to rounded, measuring on average 0.9 × 1.8 µm; sub-Stieda body is rounded to rectangular, measuring on average 1.5 × 2.6 µm; para-Stieda body is absent. Sporocyst residuum has an irregular shape consisting of numerous granules and appears membrane-bound. Sporozoites are vermiform 12.8 × 3.0 µm on average, with prominent striations at the more pointed end and two refractile bodies below striations. Segments of three gene loci (18S rRNA, 28S rRNA and cox1) were sequenced and I. lunulatae n. sp. exhibited 99.6% genetic similarity to Isospora phylidonyrisae Yang, Brice, Berto & Ryan, 2021 at the 18S rRNA gene locus, 99.8% genetic similarity to Isospora anthochaerae Yang, Brice & Ryan, 2014 and shared a 98.1% genetic similarity with Isospora manorinae Yang, Brice, Jian & Ryan, 2016 at the cox1 gene locus. Morphological and molecular data support the distinct species status of the new species.

Zoonotic infection by Cryptosporidium fayeri IVgA10G1T1R1 in a Western Australian human.

In the present study, a 37-year-old immunosuppressed female in Western Australia (WA) was identified as positive for Cryptosporidium by microscopy and treated with nitazoxanide. Molecular analyses at the 18S ribosomal RNA (18S) and 60 kDa glycoprotein (gp60) loci identified C. fayeri subtype IVgA10G1T1R1, which had previously been identified in western grey kangaroos (Macropus fuliginosus) in WA. Next generation sequencing (NGS) of the gp60 locus confirmed the absence of mixed infections with other Cryptosporidium species. This is only the second report of C. fayeri in a human host highlighting the zoonotic potential of this wildlife-associated species. Routine diagnosis using molecular methods in laboratories is required to better understand the diversity and epidemiology of Cryptosporidium parasite.