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Animal farming has intensified significantly in recent decades, with the emergence of concentrated animal feeding operations (CAFOs) in industrialized nations. The congregation of susceptible animals in CAFOs can lead to heavy environmental contamination with pathogens, promoting the emergence of hyper-transmissible, and virulent pathogens. As a result, CAFOs have been associated with emergence of highly pathogenic avian influenza viruses, hepatitis E virus, Escherichia coli O157:H7, Streptococcus suis, livestock-associated methicillin-resistant Staphylococcus aureus, and Cryptosporidium parvum in farm animals. This has led to increased transmission of zoonotic pathogens in humans and changes in disease patterns in general communities. They are exemplified by the common occurrence of outbreaks of illnesses through direct and indirect contact with farm animals, and wide occurrence of similar serotypes or subtypes in both humans and farm animals in industrialized nations. Therefore, control measures should be developed to slow down the dispersal of zoonotic pathogens associated with CAFOs and prevent the emergence of new pathogens of epidemic and pandemic potential.

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 kn

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.

The use of molecular tools has led to the identification of several zoonotic Cryptosporidium spp. in dogs and cats. Among them, Cryptosporidium canis and Cryptosporidium felis are dominant species causing canine and feline cryptosporidiosis, respectively. Some Cryptosporidium parvum infections have also been identified in both groups of animals. The identification of C. canis, C. felis and C. parvum in both pets and owners suggests the possible occurrence of zoonotic transmission of Cryptosporidium spp. between humans and pets. However, few cases of such concurrent infections have been reported. Thus, the cross-species transmission of Cryptosporidium spp. between dogs or cats and humans has long been a controversial issue. Recently developed subtyping tools for C. canis and C. felis should be very useful in identification of zoonotic transmission of both Cryptosporidium spp. Data generated using these tools have confirmed the occurrence of zoonotic transmission of these two Cryptosporidium spp. between owners and their pets, but have also shown the potential presence of host-adapted subtypes. Extensive usage of these subtyping tools in epidemiological studies of human cryptosporidiosis is needed for improved understanding of the importance of zoonotic transmission of Cryptosporidium spp. from pets.

While nutrient removal has been well studied in waste stabilisation ponds (WSPs), studies of organic micropollutant removal in pond systems are limited. In this study, we investigated organic micropollutant, nitrogen and organic carbon removal in selected WSPs that differed in geographical location and pond configuration, and compared their performance to an oxidation ditch wastewater treatment plant (WWTP). Of the 232 chemicals measured, 36 were detected in wastewater influent and 33 were detected in treated wastewater effluent. New data for micropollutant removal in WSPs was generated for three pesticides or related chemicals, five pharmaceuticals, the plasticizer N-butylbenzenesulfonamide, the antioxidant 2,6-di-t-butyl-p-cresol, and two flame retardants tris(dichloropropyl)phosphate and tris(chloropropyl)phosphate isomers. Most of these micropollutants were relatively well removed in WSPs. The poorest treatment efficiency was observed in the single facultative pond system, with no maturation pond, suggesting that the presence of a maturation pond is important for chemical removal. The two WSPs in temperate climates were found to have higher concentrations of motile algae that can optimise their position with respect to light and temperature. However, to-date, the micropollutant removal by these algal species is not known. The highest removals of micropollutants in a WSP were measured in a complex WSP system with two maturation ponds, and the removals achieved were comparable to the oxidation ditch system. The key factors contributing to high micropollutant removal in this WSP were high solar irradiation and warm temperatures that promoted the growth of non-motile green algae previously found to degrade micropollutants, and photodegradation.

Cryptosporidium spp. are common protozoan pathogens in mammals. With pet rodents being integrated into modern life, the potential roles of them in transmitting parasites to humans need assessments. In the present study, we examined the occurrence of Cryptosporidium spp. in pet rodents in Guangdong, south China. A total of 697 fecal samples were collected from 11 species of rodents in seven pet shops, one pet market and one farm. Cryptosporidium spp. were identified by PCR analysis of the small subunit rRNA gene. An overall infection rate of 36.9% (257/697) was obtained, with infection rates varying from 9.3% in chinchillas, 52.3% in guinea pigs, 57.1% in squirrels, to 68.4% in cricetid animals. Nine Cryptosporidium species and genotypes were identified, including C. wrairi (in 129 guinea pigs), C. andersoni (in 34 hamsters), C. homai (in 32 guinea pigs), Cryptosporidium hamster genotype (in 30 hamsters), C. ubiquitum (in 24 chinchillas and squirrels), C. parvum (in 2 chinchillas), Cryptosporidium ferret genotype (in 2 chipmunks), C. muris (in 1 hamster and 1 guinea pig), and Cryptosporidium chipmunk genotype V (in 1 chinchilla and 1 chipmunk). Sequence analysis of the 60 kDa glycoprotein gene identified three subtype families of C. ubiquitum, including family XIId in 15 chinchillas, XIIa in 5 chinchillas, and a new subtype family (XIIi) in 1 squirrel. The identification of C. parvum and C. ubiquitum in pet rodents suggests that these animals, especially chinchillas, could serve as reservoirs of human-pathogenic Cryptosporidium spp. Hygiene should be practiced in the rear and care of these animals, and One Health measures should be developed to reduce the occurrence of zoonotic Cryptosporidium infections due to contact with pet rodents.

Cryptosporidium is an important protozoan parasite and due to its resistance to chlorine is a major cause of swimming pool-associated gastroenteritis outbreaks. The present study combined contact tracing and molecular techniques to analyse cryptosporidiosis cases and outbreaks in Western Australia in 2019 and 2020. In the 2019 outbreak, subtyping at the 60 kDa glycoprotein (gp60) gene identified 89.0% (16/18) of samples were caused by the C. hominis IdA15G1 subtype. Amplicon next generation sequencing (NGS) at the gp60 locus identified five C. hominis IdA15G1 subtype samples that also had C. hominis IdA14 subtype DNA, while multi locus sequence typing (MLST) analysis on a subset (n = 14) of C. hominis samples identified three IdA15G1 samples with a 6 bp insertion at the end of the trinucleotide repeat region of the cp47 gene. In 2020, 88.0% (73/83) of samples typed were caused by the relatively rare C. hominis subtype IbA12G3. Four mixed infections were observed by NGS with three IdA15G1/ IdA14 mixtures and one C. parvum IIaA18G3R1 sample mixed with IIaA16G3R1. No genetic diversity using MLST was detected. Epidemiological and molecular data indicates that the outbreaks in 2019 and 2020 were each potentially from swimming pool point sources and a new C. hominis subtype IbA12G3 is emerging in Australia. The findings of the present study are important for understanding the introduction and transmission of rare Cryptosporidium subtypes to vulnerable populations.

A new Isospora species is described from New Holland honeyeaters (Phylidonyris novaehollandiae). Sporulated oocysts (n = 25) were characterised as subspheroidal, 29–32 × 28–31 (29.8 × 29.4); length/width (L/W) ratio 1.01–1.02 (1.01). Wall bi-layered, 1.3–1.6 (1.5) thick, outer layer smooth, c.2/3 of total thickness. Micropyle and oocyst residuum absent, but usually two polar granules are present. Sporocysts (n = 25) ovoidal, 18–19 × 12–14 (18.4 × 12.3); L/W ratio 1.42–1.53 (1.50). Stieda body present, flattened, c.0.5 deep × 2.5 wide; sub-Stieda present, rounded, c.2.5 deep × 3.5 wide; para-Stieda body absent; sporocyst residuum present, usually a distinctly irregular body consisting of numerous small granules that appear to be membrane-bound. Sporozoites vermiform, with robust anterior and posterior refractile bodies. Molecular characterization was conducted at the 18S and 28S ribosomal RNA and the mitochondrial (mt) cytochrome oxidase (COI) loci. Phylogenetic analysis of genomic 18S and mt COI sequences indicated that Isospora phylidonyrisae n. sp. was genetically similar to Isospora coronoideae, isolated from an Australian raven (Corvus coronoides) in Western Australia, with a 99.3% and 98.4% homology, respectively. The 28S rRNA sequence was most similar to Isospora anthochaerae (KF766053) and Isospora manorinae (KT224381), both with a 98.2% genetic similarity. Based on morphological and genetic data, this isolate is a new species of Isospora, which is named Isospora phylidonyrisae n. sp. after its host.

Sheep and goats are commonly infected with three Cryptosporidium species, including Cryptosporidium parvum, Cryptosporidium ubiquitum, and Cryptosporidium xiaoi, which differ from each in prevalence, geographic distribution, and public health importance. While C. parvum appears to be a dominant species in small ruminants in European countries, its occurrence in most African, Asian, and American countries appear to be limited. As a result, zoonotic infections due to contact with lambs and goat kids are common in European countries, leading to frequent reports of outbreaks of cryptosporidiosis on petting farms. In contrast, C. xiaoi is the dominant species elsewhere, and mostly does not infect humans. While C. ubiquitum is another zoonotic species, it occurs in sheep and goats at much lower frequency. Host adaptation appears to be present in both C. parvum and C. ubiquitum, consisting of several subtype families with different host preference. The host-adapted nature of C. parvum and C. ubiquitum has allowed the use of subtyping tools in tracking infection sources. This has led to the identification of geographic differences in the importance of small ruminants in epidemiology of human cryptosporidiosis. These tools have also been used effectively in linking zoonotic transmission of C. parvum between outbreak cases and the suspected animals. Further studies should be directly elucidating the reasons for differences in the distribution and public health importance of major Cryptosporidium species in sheep and goats.

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.

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.

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.

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.

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.

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.

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.

Giardia duodenalis is a common intestinal parasite in various hosts, with the disease giardiasis being a zoonosis. The use of molecular typing tools has improved our understanding of the distribution and zoonotic potential of G. duodenalis genotypes in different animals. The present review summarizes recent data on the distribution of G. duodenalis genotypes in humans and animals in different areas. The dominance of G. duodenalis assemblages A and B in humans and common occurrence of host-adapted assemblages in most domesticated animals suggests that zoonotic giardiasis is probably less common than believed and could be attributed mainly to contact with or contamination from just a few species of animals such as nonhuman primates, equines, rabbits, guinea pigs, chinchillas, and beavers. Future studies should be directed to advanced genetic characterization of isolates from well-designed epidemiological investigations, especially comparative analyses of isolates from humans and animals living in the same household or community. This will likely lead to better understanding of zoonotic transmission of G. duodenalis in different environmental and socioeconomic settings.

Current knowledge of Cryptosporidium species/genotypes in marine fish is limited. Following phylogenetic analysis at the 18S rDNA locus, a recent study identified six new genotypes of Cryptosporidium colonizing edible fish found in European seas. Of these, five grouped in a clade together (#Cryptofish 1–5) and one grouped separately (#Cryptofish 7). In the present study, after phylogenetic analyses of #Cryptofish1, #Cryptofish2, #Cryptofish4, #Cryptofish5 and #Cryptofish7 at the actin locus, the presence of two major clades was confirmed. In addition, when possible, longer 18S amplicons were generated. In conclusion, the small genetic distances between these genotypes designated as a novel marine genotype I (#Cryptofish 1-5) suggest that they may be genetic variants of the same species, while the designated novel marine genotype 2 (#Cryptofish 7) is clearly representative of a separate species.

Cryptosporidium and Giardia infections can negatively impact livestock health and reduce productivity, and some species and genotypes infecting livestock have zoonotic potential. Infection occurs via the faecal-oral route. Waterborne infections are a recognised source of infection for humans, but the role of livestock drinking water as a source of infection in livestock has not been described. This study aimed to determine whether contaminated drinking water supplies, such as farm dams, are a likely transmission source for Cryptosporidium and Giardia infections for extensively managed sheep. Dam water samples (n = 47) were collected during autumn, winter and spring from 12 farm dams located on six different farms in south west Western Australia, and faecal samples (n = 349) were collected from sheep with access to these dams. All samples were initially screened for Cryptosporidium spp. at the 18S locus and Giardia spp. at the gdh gene using qPCR, and oocyst numbers were determined directly from the qPCR data using DNA standards calibrated by droplet digital PCR. Cryptosporidium-positive sheep faecal samples were typed and subtyped by sequence analysis of 18S and gp60 loci, respectively. Giardia-specific PCR and Sanger sequencing targeting tpi and gdh loci were performed on Giardia- positive sheep faecal samples to characterise Giardia duodenalis assemblages. To identify Cryptosporidium and Giardia spp. in dam water samples, next-generation sequencing analysis of 18S and gdh amplicons were performed, respectively. Two species of Cryptosporidium (Cryptosporidium xiaoi and Cryptospordium ubiquitum (subtype family XIIa)) were detected in 38/345 sheep faecal samples, and in water from 9/12 farm dams during the study period, with C. xiaoi the species most frequently detected in both faeces and dam water overall. Giardia duodenalis assemblages AI, AII and E were detected in 36/348 faecal samples and water from 10/12 farm dams. For dam water samples where oo/cysts were dete

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.