Use of micropollutant indicator ratios to characterize wastewater treatment plant efficiency and to identify wastewater impact on groundwater.

Contamination by wastewater has been traditionally assessed by measuring faecal coliforms, such as E. coli and entereococci. However, using micropollutants to track wastewater input is gaining interest. In this study, we identified nine micropollutant indicators that could be used to characterize water quality and wastewater treatment efficiency in pond-based wastewater treatment plants (WWTPs) of varying configuration. Of 232 micropollutants tested, nine micropollutants were detected in treated wastewater at concentrations and frequencies suitable to be considered as indicators for treated wastewater. The nine indicators were then classified as stable (carbamazepine, sucralose, benzotriazole, 4+5-methylbenzotriazole), labile (atorvastatin, naproxen, galaxolide) or intermediate/uncertain (gemfibrozil, tris(chloropropyl)phosphate isomers) based on observed removals in the pond-based WWTPs and correlations between micropollutant and dissolved organic carbon removal. The utility of the selected indicators was evaluated by assessing the wastewater quality in different stages of wastewater treatment in three pond-based WWTPs, as well as selected groundwater bores near one WWTP, where treated wastewater was used to irrigate a nearby golf course. Ratios of labile to stable indicators provided insight into the treatment efficiency of different facultative and maturation ponds and highlighted the seasonal variability in treatment efficiency for some pond-based WWTPs. Additionally, indicator ratios of labile to stable indicators identified potential unintended release of untreated wastewater to groundwater, even with the presence of micropollutants in other groundwater bores related to approved reuse of treated wastewater.

Minimal zoonotic risk of cryptosporidiosis and giardiasis from frogs and reptiles.

The zoonotic potential of the protist parasites Cryptosporidium spp. and Giardia duodenalis in amphibians and reptiles raises public health concerns due to their growing popularity as pets. This review examines the prevalence and diversity of these parasites in wild and captive amphibians and reptiles to better understand the zoonotic risk. Research on Giardia in both groups is limited, and zoonotic forms of Cryptosporidium or Giardia have not been reported in amphibians. Host-adapted Cryptosporidium species dominate in reptiles, albeit some reptiles have been found to carry zoonotic (C. hominis and C. parvum) and rodent-associated (C. tyzzeri, C. muris and C. andersoni) species, primarily through mechanical carriage. Similarly, the limited reports of Giardia duodenalis (assemblages A, B and E) in reptiles may also be due to mechanical carriage. Thus, the available evidence indicates minimal zoonotic risk associated with these organisms in wild and captive frogs and reptiles. The exact transmission routes for these infections within reptile populations remain poorly understood, particularly regarding the importance of mechanical carriage. Although the risk appears minimal, continued research and surveillance efforts are necessary to gain a more comprehensive understanding of the transmission dynamics and ultimately improve our ability to safeguard human and animal health.

Rabbits as reservoirs: An updated perspective of the zoonotic risk from Cryptosporidium and Giardia.

Rabbits are highly abundant in many countries and can serve as reservoirs of diseases for a diversity of pathogens including the enteric protozoan parasites, Cryptosporidium and Giardia. Both parasites shed environmentally robust environmental stages (oo/cysts) and have been responsible for numerous waterborne outbreaks of diseases. Cryptosporidium hominis and C. parvum are responsible for most infections in humans, while Giardia duodenalis assemblages A and B, cause most human cases of giardiasis. Cryptosporidium cuniculus, the dominant species infecting rabbits, is the only spceies other than C. hominis and C. parvum to have caused a waterborne outbreak of gastritis, which occurred in the United Kingdom in 2008. This review examines the prevalence of Cryptosporidium and Giardia species in rabbits to better understand the public health risks of contamination of water sources with Cryptosporidium and Giardia oo/cysts from rabbits. Despite the abundance of C. cuniculus in rabbits, reports in humans are relatively rare, with the exception of the United Kingdom and New Zealand, and reports of C. cuniculus in humans from the United Kingdom have declined substantially since the 2008 outbreak. Subtyping of C. cuniculus has supported the potential for zoonotic transmission. Relatively few studies have been conducted on Giardia, but assemblage B dominates. However, improved typing methods are required to better understand the transmission dynamics of Giardia assemblages in rabbits. Similarly, it is not well understood if pet rabbits or contaminated water are the main source of C. cuniculus infections in humans. Well-planned studies using high-resolution typing tools are required to understand the transmission dynamics better and quantify the public health risk of Cryptosporidium and Giardia from rabbits.

Critters and contamination: Zoonotic protozoans in urban rodents and water quality.

Rodents represent the single largest group within mammals and host a diverse array of zoonotic pathogens. Urbanisation impacts wild mammals, including rodents, leading to habitat loss but also providing new resources. Urban-adapted (synanthropic) rodents, such as the brown rat (R. norvegicus), black rat (R. rattus), and house mouse (Mus musculus), have long successfully adapted to living close to humans and are known carriers of zoonotic pathogens. Two important enteric, zoonotic protozoan parasites, carried by rodents, include Cryptosporidium and Giardia. Their environmental stages (oocysts/cysts), released in faeces, can contaminate surface and wastewaters, are resistant to common drinking water disinfectants and can cause water-borne related gastritis outbreaks. At least 48 species of Cryptosporidium have been described, with C. hominis and C. parvum responsible for the majority of human infections, while Giardia duodenalis assemblages A and B are the main human-infectious assemblages. Molecular characterisation is crucial to assess the public health risk linked to rodent-related water contamination due to morphological overlap between species. This review explores the global molecular diversity of these parasites in rodents, with a focus on evaluating the zoonotic risk from contamination of water and wasterwater with Cryptosporidium and Giardia oocysts/cysts from synanthropic rodents. Analysis indicates that while zoonotic Cryptosporidium and Giardia are prevalent in farmed and pet rodents, host-specific Cryptosporidium and Giardia species dominate in urban adapted rodents, and therefore the risks posed by these rodents in the transmission of zoonotic Cryptosporidium and Giardia are relatively low. Many knowledge gaps remain however, and therefore understanding the intricate dynamics of these parasites in rodent populations is essential for managing their impact on human health and water quality. This knowledge can inform strategies to reduce disease transmission and ensure safe drinking water in urban and peri­urban areas.

Zoonotic Cryptosporidium and Giardia in marsupials-an update.

Marsupials, inhabiting diverse ecosystems, including urban and peri-urban regions in Australasia and the Americas, intersect with human activities, leading to zoonotic spill-over and anthroponotic spill-back of pathogens, including Cryptosporidium and Giardia. This review assesses the current knowledge on the diversity of Cryptosporidium and Giardia species in marsupials, focusing on the potential zoonotic risks. Cryptosporidium fayeri and C. macropodum are the dominant species in marsupials, while in possums, the host-specific possum genotype dominates. Of these three species/genotypes, only C. fayeri has been identified in two humans and the zoonotic risk is considered low. Generally, oocyst shedding in marsupials is low, further supporting a low transmission risk. However, there is some evidence of spill-back of C. hominis into kangaroo populations, which requires continued monitoring. Although C. hominis does not appear to be established in small marsupials like possums, comprehensive screening and analysis are essential for a better understanding of the prevalence and potential establishment of zoonotic Cryptosporidium species in small marsupials. Both host-specific and zoonotic Giardia species have been identified in marsupials. The dominance of zoonotic G. duodenalis assemblages A and B in marsupials may result from spill-back from livestock and humans and it is not yet understood if these are transient or established infections. Future studies using multilocus typing tools and whole-genome sequencing are required for a better understanding of the zoonotic risk from Giardia infections in marsupials. Moreover, much more extensive screening of a wider range of marsupial species, particularly in peri-urban areas, is required to provide a clearer understanding of the zoonotic risk of Cryptosporidium and Giardia in marsupials.

The risk of wild birds contaminating source water with zoonotic Cryptosporidium and Giardia is probably overestimated.

Cryptosporidium and Giardia are important waterborne protozoan parasites that are resistant to disinfectants commonly used for drinking water. Wild birds, especially wild migratory birds, are often implicated in the contamination of source and wastewater with zoonotic diseases, due to their abundance near water and in urban areas and their ability to spread enteric pathogens over long distances. This review summarises the diversity of Cryptosporidium and Giardia in birds, with a focus on zoonotic species, particularly in wild and migratory birds, which is critical for understanding zoonotic risks. The analysis revealed that both avian-adapted and zoonotic Cryptosporidium species have been identified in birds but that avian-adapted Cryptosporidium species dominate in wild migratory birds. Few studies have examined Giardia species and assemblages in birds, but the non-zoonotic Giardia psittaci and Giardia ardeae are the most commonly reported species. The identification of zoonotic Cryptosporidium and Giardia in birds, particularly C. parvum and G. duodenalis assemblages A and B in wild migratory birds, is likely due to mechanical carriage or spillback from birds co-grazing pastures contaminated with C. parvum from livestock. Therefore, the role of wild migratory birds in the transmission of zoonotic Cryptosporidium and Giardia to source water is likely overestimated. To address knowledge gaps, it is important to conduct more extensive studies on the prevalence of Cryptosporidium and Giardia in a broader range of migratory wild birds. There is also a need to investigate the extent to which zoonotic infections with C. hominis/C. parvum and G. duodenalis assemblages A and B are mechanical and/or transient, and to assess the load and viability of zoonotic oo/cysts shed in avian faeces. Understanding the contribution of birds to zoonoses is essential for effective disease surveillance, prevention, and control.

Cryptosporidium and Giardia in cats and dogs: What is the real zoonotic risk?

Due to the close bond between humans and companion animals, a thorough understanding of the diversity of Cryptosporidium species and Giardia assemblages in cats and dogs is essential to determine the potential zoonotic risks. Analysis of molecular studies shows that C. felis and C. canis are the main species infecting cats and dogs, respectively. These species are largely host-specific, as despite intense association with humans, prevalence of C. felis and C. canis in humans is low and predominantly in immunocompromised individuals and low-income countries. There have been reports of C. parvum in cats and dogs and two reports of C. hominis in dogs. In most studies conducted to date, however, the prevalence of zoonotic species was low and may be associated with coprophagy and or/spillback, but this remains to be determined. Results of subtyping studies suggest that for C. felis and C. canis, some zoonotic transmission may occur but host-adapted subtypes also exist. Giardia duodenalis assemblages C and D are commonly reported in dogs, with assemblages F and A most common in cats. Assemblages C, D and F are largely host-specific as there are only a handful of reports of them in humans. Reports of assemblage A and B in cats and dogs may be due to coprophagy or spillback from owners. Despite the extent of pet ownership and the close contact between humans and companion animals worldwide, the overall risk of zoonotic transmission from cats and dogs to humans is uncertain but thought to be low due to C. canis, C. felis and G. duodenalis assemblages C, D and F being predominantly host-specific, the relatively low prevalence of C. parvum (and C. hominis) in cats and dogs (which may be due to mechanical carriage), and low oo/cyst shedding. Carefully designed epidemiological studies of cats and dogs and their owners using subtyping tools are essential to better quantify the extent of spillover and spillback of Cryptosporidium and Giardia between pets and their owners.

How significant are bats as potential carriers of zoonotic Cryptosporidium and Giardia?

Bats are known to harbour various pathogens and are increasingly recognised as potential reservoirs for zoonotic diseases. This paper reviews the genetic diversity and zoonotic potential of Cryptosporidium and Giardia in bats. The risk of zoonotic transmission of Cryptosporidium from bats to humans appears low, with bat-specific Cryptosporidium genotypes accounting for 91.5% of Cryptosporidium-positive samples genotyped from bats worldwide, and C. parvum and C. hominis accounting for 3.4% each of typed positives, respectively. To date, there have only been sporadic detections of Giardia in bats, with no genetic characterisation of the parasite to species or assemblage level. Therefore, the role bats play as reservoirs of zoonotic Giardia spp. is unknown. To mitigate potential risks of zoonotic transmission and their public health implications, comprehensive research on Cryptosporidium and Giardia in bats is imperative. Future studies should encompass additional locations across the globe and a broader spectrum of bat species, with a focus on those adapted to urban environments.

Comparison of in vitro growth characteristics of Cryptosporidium hominis (IdA15G1) and Cryptosporidium parvum (Iowa-IIaA17G2R1 and IIaA18G3R1).

Cryptosporidium is a major cause of diarrhoeal disease and mortality in young children in resource-poor countries, for which no vaccines or adequate therapeutic options are available. Infection in humans is primarily caused by two species: C. hominis and C. parvum. Despite C. hominis being the dominant species infecting humans in most countries, very little is known about its growth characteristics and life cycle in vitro, given that the majority of our knowledge of the in vitro development of Cryptosporidium has been based on C. parvum. In the present study, the growth and development of two C. parvum isolates (subtypes Iowa-IIaA17G2R1 and IIaA18G3R1) and one C. hominis isolate (subtype IdA15G1) in HCT-8 cells were examined and compared at 24 h and 48 h using morphological data acquired with scanning electron microscopy. Our data indicated no significant differences in the proportion of meronts or merozoites between species or subtypes at either time-point. Sexual development was observed at the 48-h time-point across both species through observations of both microgamonts and macrogamonts, with a higher frequency of macrogamont observations in C. hominis (IdA15G1) cultures at 48-h post-infection compared to both C. parvum subtypes. This corresponded to differences in the proportion of trophozoites observed at the same time point. No differences in proportion of microgamonts were observed between the three subtypes, which were rarely observed across all cultures. In summary, our data indicate that asexual development of C. hominis is similar to that of C. parvum, while sexual development is accelerated in C. hominis. This study provides new insights into differences in the in vitro growth characteristics of C. hominis when compared to C. parvum, which will facilitate our understanding of the sexual development of both species.

Longitudinal follow-up reveals occurrence of successive Cryptosporidium bovis and Cryptosporidium ryanae infections by different subtype families in dairy cattle.

Cryptosporidium bovis and Cryptosporidium ryanae are common species causing cryptosporidiosis in cattle. Data accumulated thus far indicate that the infection patterns of the two species could be different between areas with and without Cryptosporidium parvum. To better understand the infection dynamics of these two species, cross-sectional and longitudinal studies of Cryptosporidium spp. were conducted using genotyping and subtyping tools. In the cross-sectional survey, analysis of 634 faecal samples from two farms identified only C. bovis and C. ryanae in pre-weaned calves. Two birth cohorts of 61 and 78 calves were followed longitudinally over a 12 month period, which revealed the shedding of C. bovis oocysts started at 1-2 weeks of age and peaked initially at 6-8 weeks of age. Altogether calves experienced four infections by six subtype families of C. bovis, with each infection caused by different subtype families. In contrast, the shedding of C. ryanae oocysts started at 2-4 weeks of age, and the two infections were caused by different subtype families. The cumulative incidence of C. bovis infection was 100% (58/58, 32/32) on both farms, compared with 84.4-98.3% (27/32 and 57/58) for C. ryanae infection. Overall, the mean duration of oocyst shedding in the cohort studies was 3.8-4.0 weeks for C. bovis compared with 2.1 weeks for C. ryanae. The oocyst shedding intensity was high (mean oocysts per gram of faeces was over 105) during the first infection with each species but became significantly lower in the later infections. Cryptosporidium ryanae was associated with the occurrence of diarrhea on one farm, while C. bovis was not. The data indicate that there is an early occurrence of C. bovis and C. ryanae in pre-weaned calves with high infection intensity in the absence of C. parvum. Calves infected with the same Cryptosporidium sp. multiple times could be associated with the presence of subtype-specific immunity.

Cryptosporidium equi n. sp. (Apicomplexa: Cryptosporidiidae): Biological and genetic characterisations.

The horse genotype is one of three common Cryptosporidium spp. in equine animals and has been identified in some human cases. The species status of Cryptosporidium horse genotype remains unclear due to the lack of extensive morphological, biological, and genetic data. In the present study, we have conducted biological and whole genome sequence analyses of an isolate of the genotype from hedgehogs and proposed to name it Cryptosporidium equi n. sp. to reflect its common occurrence in equine animals. Oocysts of C. equi measured 5.12 ± 0.36 µm × 4.46 ± 0.21 µm with a shape index of 1.15 ± 0.08 (n = 50). Cryptosporidium equi was infectious to 3-week-old four-toed hedgehogs (Atelerix albiventris) and mice, with a prepatent period of 2-9 days and a patent period of 30-40 days in hedgehogs. It was not infectious to rats and rabbits. Phylogenetic analyses of small subunit rRNA, 70 kDa heat shock protein, actin, 60 kDa glycoprotein and 100 other orthologous genes revealed that C. equi is genetically distinct from other known Cryptosporidium species and genotypes. The sequence identity between C. equi and Cryptosporidium parvum genomes is 97.9%. Compared with C. parvum, C. equi has lost two MEDLE genes and one insulinase-like protease gene and gained one SKSR gene. In addition, 60 genes have highly divergent sequences (sequence differences ≥ 5.0%), including those encoding mucin-like glycoproteins, insulinase-like peptidases, and MEDLE and SKSR proteins. The genetic uniqueness of C. equi supports its increasing host range and the naming of it as a valid Cryptosporidium species. This is the first known use of whole genome sequence data in delineating new Cryptosporidium species.

Detection of unusual Cryptosporidium parvum subtype in patients with gastrointestinal cancer in Egypt.

While the importance of cryptosporidiosis in immunocompromised persons is well known, the prevalence of Cryptosporidium spp. in cancer patients is not clear. The current study was designed to assess the occurrence and genetic characteristics of Cryptosporidium spp. in patients with gastrointestinal (GI) cancer in Egypt. Stool samples were collected from 100 patients with GI malignancies and 20 healthy individuals without any GI manifestations (control group). They were screened by microscopy and the immunochromatographic RIDA®QUICK Cryptosporidium kit. Subtyping of Cryptosporidium spp. was conducted by sequence analysis of the glycoprotein 60 (gp60) locus. Sociodemographic, environmental data and information on GI symptoms, cancer types, and clinical treatment were obtained via a questionnaire. By microscopy and RIDA®QUICK, only 7% (7/100) of GI cancer patients were positive for Cryptosporidium, compared with 40% (40/100) by gp60 nPCR. No positives were obtained from the control group. Male sex (P = 0.02) and younger age (P = 0.004) were major Cryptosporidium risk factors for infection. The occurrence of Cryptosporidium was also significantly more frequent (P = 0.003) in watery stool samples. Sequence analysis of the gp60 amplicons (~ 400 bp) identified a novel C. parvum subtype with nine TCA repeats and eleven ACATCA repeats. A formal subtype designation could not be made due to the short sequence length. More studies should be conducted to verify the common occurrence of this unusual C. parvum subtype and establish its genetic identity.

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.

Point of care diagnostics for Cryptosporidium: new and emerging technologies.

PURPOSE OF REVIEW: Although Cryptosporidium detection and typing techniques have improved dramatically in recent years, relatively little research has been conducted on point of care (POC) detection and typing tools. Therefore, the main purpose of the present review is to summarize and evaluate recent and emerging POC diagnostic methods for Cryptosporidium spp. RECENT FINDINGS: Microscopy techniques such as light-emitting diode fluorescence microscopy with auramine-phenol staining (LED-AP), still have utility for (POC) diagnostics but require fluorescent microscopes and along with immunological-based techniques, suffer from lack of specificity and sensitivity. Molecular detection and typing tools offer higher sensitivity, specificity and speciation, but are currently too expensive for routine POC diagnostics. Isothermal amplification methods such as loop-mediated isothermal amplification (LAMP) or recombinase polymerase amplification (RPA) including a commercially available LAMP kit have been developed for Cryptosporidium but are prone to false positives. Clustered regularly interspaced short palindromic repeats (CRISPR)-Cas diagnostic technologies (CRISPRDx) have recently been combined with isothermal amplification to increase its specificity and sensitivity for detection and typing. Other emerging technologies including amplification-free CRISPR detection methods are currently being developed for Cryptosporidium using a smartphone to read the results. SUMMARY: Many challenges are still exist in the development of POC diagnostics for Cryptosporidium. The ideal POC tool would be able to concentrate the pathogen prior to detection and typing, which is complicated and research in this area is still very limited. In the short-term, CRISPR-powered isothermal amplification lateral flow tools offer the best opportunity for POC Cryptosporidium species and subtype detection, with a fully integrated autonomous biosensor for the long-term goal.

The Troublesome Ticks Research Protocol: Developing a Comprehensive, Multidiscipline Research Plan for Investigating Human Tick-Associated Disease in Australia.

In Australia, there is a paucity of data about the extent and impact of zoonotic tick-related illnesses. Even less is understood about a multifaceted illness referred to as Debilitating Symptom Complexes Attributed to Ticks (DSCATT). Here, we describe a research plan for investigating the aetiology, pathophysiology, and clinical outcomes of human tick-associated disease in Australia. Our approach focuses on the transmission of potential pathogens and the immunological responses of the patient after a tick bite. The protocol is strengthened by prospective data collection, the recruitment of two external matched control groups, and sophisticated integrative data analysis which, collectively, will allow the robust demonstration of associations between a tick bite and the development of clinical and pathological abnormalities. Various laboratory analyses are performed including metagenomics to investigate the potential transmission of bacteria, protozoa and/or viruses during tick bite. In addition, multi-omics technology is applied to investigate links between host immune responses and potential infectious and non-infectious disease causations. Psychometric profiling is also used to investigate whether psychological attributes influence symptom development. This research will fill important knowledge gaps about tick-borne diseases. Ultimately, we hope the results will promote improved diagnostic outcomes, and inform the safe management and treatment of patients bitten by ticks in Australia.

Current assumptions for quantitative microbial risk assessment (QMRA) of Norovirus contamination of drinking water catchments due to recreational activities: an update.

Contamination of drinking water from Norovirus (NoV) and other waterborne viruses is a major public health concern globally. Increasingly, quantitative microbial risk assessment (QMRA) is being used to assess the various risks from waterborne pathogens and evaluate control strategies. As urban populations grow and expand, there is increasing demand for recreational activities in drinking water catchments. QMRA relies on context-specific data to map out the pathways by which viruses can enter water and be transferred to drinking water consumers and identify risk factors and appropriate controls. This review examines the current evidence base and assumptions for QMRA analysis of NoV and other waterborne viral pathogens and recommends numerical values based on the most recent evidence to better understand the health risks associated with recreators in Australian drinking water sources; these are broadly applicable to all drinking water sources where recreational access is allowed. Key issues include the lack of an agreed upon data and dose-response models for human infectious NoV genotypes, faecal shedding by bathers, the extent of NoV infectivity and aggregation, resistance (secretor status) to NoV and the extent of secondary transmission.

Emergence of zoonotic Cryptosporidium parvum in China.

Zoonotic cryptosporidiosis is a major public health problem in industrialized nations; in those countries it is caused mainly by Cryptosporidium parvum IIa subtypes that are prevalent in dairy calves. Because of the short history of intensive animal farming in China, strains of C. parvum are found only on some dairy farms in this country and are the IId subtypes. However, the prevalence of C. parvum is increasing rapidly, with IIa subtypes recently detected in a few grazing animals, and both IIa and IId subtypes are emerging in humans. As animal farming intensifies, China may follow in the footsteps of industrialized nations where zoonotic cryptosporidiosis is rampant. One Health and biosecurity measures are urgently needed to slow down the dispersal of autochthonous IId subtypes and imported IIa subtypes.

A review of the molecular epidemiology of Cryptosporidium spp. and Giardia duodenalis in the Middle East and North Africa (MENA) region.

Cryptosporidium spp. and Giardia duodenalis are important protozoan parasites which are associated with diarrheal diseases in humans and animals worldwide. Relatively little is known about the molecular epidemiology of Cryptosporidium spp. and Giardia duodenalis in the Middle East Countries and North Africa (MENA region). Therefore, this review aimed to inspect published genotyping and subtyping studies on Cryptosporidium spp. and Giardia duodenalis in the MENA region. These studies indicate that both anthroponotic and zoonotic transmission of Cryptosporidium occurs with the predominance of zoonotic transmission in most countries. Seven Cryptosporidium species were identified in humans (C. parvum, C. hominis, Cryptosporidium meleagridis, C. felis, Cryptosporidium muris, C. canis and C. bovis), with C. parvum by far being the most prevalent species (reported in 95.4% of the retrieved studies). Among C. parvum gp60 subtype families, IIa and IId predominated, suggesting potential zoonotic transmission. However, in four MENA countries (Lebanon, Israel, Egypt and Tunisia), C. hominis was the predominant species with five subtype families reported including Ia, Ib, Id, If and Ie, all of which are usually anthroponotically transmitted between humans. In animals, the majority of studies were conducted mainly on livestock and poultry, 15 species were identified (C. parvum, C. hominis, C. muris, Cryptosporidium cuniculus, C. andersoni, C. bovis, C. meleagridis, C. baileyi, C. erinacei, C. ryanae, C. felis, C. suis, Cryptosporidium galli, C. xiaoi and C. ubiquitum) with C. parvum (IIa and IId subtypes) the dominant species in livestock and C. meleagridis and C. baileyi the dominant species in poultry. With G. duodenalis, five assemblages (A, B, C, E and F) were identified in humans and six (A, B, C, E, D and F) in animals in MENA countries with assemblages A and B commonly reported in humans, and assemblages A and E dominant in livestock. This review also identified a major knowledge gap in the lack of Cryptosporidium spp. and Giardia duodenalis typing studies in water and food sources in the MENA region. Of the few studies conducted on water sources (including drinking and tap water), ten Cryptosporidium species and four genotypes were identified, highlighting the potential role of water as the major route of Cryptosporidium spp. transmission in the region. In addition, three G. duodenalis assemblages (A, B and E) were detected in different water sources with AI, AII and BIV being the main sub-assemblages reported. More research is required in order to better understand the molecular diversity and transmission dynamics of Cryptsporidum spp. and Giardia duodenalis in humans, animals, water and food sources in MENA region.

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.

The bacterial biome of ticks and their wildlife hosts at the urban-wildland interface.

Advances in sequencing technologies have revealed the complex and diverse microbial communities present in ticks (Ixodida). As obligate blood-feeding arthropods, ticks are responsible for a number of infectious diseases that can affect humans, livestock, domestic animals and wildlife. While cases of human tick-borne diseases continue to increase in the northern hemisphere, there has been relatively little recognition of zoonotic tick-borne pathogens in Australia. Over the past 5 years, studies using high-throughput sequencing technologies have shown that Australian ticks harbour unique and diverse bacterial communities. In the present study, free-ranging wildlife (n=203), representing ten mammal species, were sampled from urban and peri-urban areas in New South Wales (NSW), Queensland (QLD) and Western Australia (WA). Bacterial metabarcoding targeting the 16S rRNA locus was used to characterize the microbiomes of three sample types collected from wildlife: blood, ticks and tissue samples. Further sequence information was obtained for selected taxa of interest. Six tick species were identified from wildlife: Amblyomma triguttatum, Ixodes antechini, Ixodes australiensis, Ixodes holocyclus, Ixodes tasmani and Ixodes trichosuri. Bacterial 16S rRNA metabarcoding was performed on 536 samples and 65 controls, generating over 100 million sequences. Alpha diversity was significantly different between the three sample types, with tissue samples displaying the highest alpha diversity (P<0.001). Proteobacteria was the most abundant taxon identified across all sample types (37.3 %). Beta diversity analysis and ordination revealed little overlap between the three sample types (P<0.001). Taxa of interest included Anaplasmataceae, Bartonella, Borrelia, Coxiellaceae, Francisella, Midichloria, Mycoplasma and Rickettsia. Anaplasmataceae bacteria were detected in 17.7% (95/536) of samples and included Anaplasma, Ehrlichia and Neoehrlichia species. In samples from NSW, 'Ca. Neoehrlichia australis', 'Ca. Neoehrlichia arcana', Neoehrlichia sp. and Ehrlichia sp. were identified. A putative novel Ehrlichia sp. was identified from WA and Anaplasma platys was identified from QLD. Nine rodent tissue samples were positive for a novel Borrelia sp. that formed a phylogenetically distinct clade separate from the Lyme Borrelia and relapsing fever groups. This novel clade included recently identified rodent-associated Borrelia genotypes, which were described from Spain and North America. Bartonella was identified in 12.9% (69/536) of samples. Over half of these positive samples were obtained from black rats (Rattus rattus), and the dominant bacterial species identified were Bartonella coopersplainsensis and Bartonella queenslandensis. The results from the present study show the value of using unbiased high-throughput sequencing applied to samples collected from wildlife. In addition to understanding the sylvatic cycle of known vector-associated pathogens, surveillance work is important to ensure preparedness for potential zoonotic spillover events.