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Giardia is a prevalent single-celled microaerophilic intestinal parasite causing diarrheal disease and significantly impacting global health. Double diploid (essentially tetraploid) Giardia trophozoites have presented a formidable challenge to the development of molecular genetic tools to interrogate gene function. High sequence divergence and the high percentage of hypothetical proteins lacking homology to proteins in other eukaryotes have limited our understanding of Giardia protein function, slowing drug target validation and development. For more than 25 years, Giardia A and B assemblages have been readily amenable to transfection with plasmids or linear DNA templates. Here, we highlight the utility and power of genetic approaches developed to assess protein function in Giardia, with particular emphasis on the more recent clustered regularly interspaced palindromic repeats/Cas9-based methods for knockdowns and knockouts. Robust and reliable molecular genetic approaches are fundamental toward the interrogation of Giardia protein function and evaluation of druggable targets. New genetic approaches tailored for the double diploid Giardia are imperative for understanding Giardia's unique biology and pathogenesis.

The intestinal parasites Giardia lamblia and Entamoeba histolytica are major causes of morbidity and mortality associated with diarrheal diseases. Metronidazole is the most common drug used to treat giardiasis and amebiasis. Despite its efficacy, treatment failures in giardiasis occur in up to 5%-40% of cases. Potential resistance of E. histolytica to metronidazole is an increasing concern. Therefore, it is critical to search for more effective drugs to treat giardiasis and amebiasis. We identified antigiardial and antiamebic activities of the rediscovered nitroimidazole compound, fexinidazole, and its sulfone and sulfoxide metabolites. Fexinidazole is equally active against E. histolytica and G. lamblia trophozoites, and both metabolites were 3- to 18-fold more active than the parent drug. Fexinidazole and its metabolites were also active against a metronidazole-resistant strain of G. lamblia. G. lamblia and E. histolytica cell extracts exhibited decreased residual nitroreductase activity when metabolites were used as substrates, indicating nitroreductase may be central to the mechanism of action of fexinidazole. In a cell invasion model, fexinidazole and its metabolites significantly reduced the invasiveness of E. histolytica trophozoites through basement membrane matrix. A q.d. oral dose of fexinidazole and its metabolites at 10 mg/kg for 3 days reduced G. lamblia infection significantly in mice compared to control. The newly discovered antigiardial and antiamebic activities of fexinidazole, combined with its FDA-approval and inclusion in the WHO Model List of Essential Medicines for the treatment of human African trypanosomiasis, offer decreased risk and a shortened development timeline toward clinical use of fexinidazole for treatment of giardiasis or amebiasis.

This review of literature analyses the effectiveness of three commercially available filters, namely the Pall Envirochek filters, IDEXX FiltaMax, and Whatman flatbed membrane, in the process of concentrating Cryptosporidium oocysts and Giardia cysts from water samples. Moreover, several research studies investigated the possibility of using ultrafiltration filters as an affordable substitute for pathogen concentration, and their efficacy for protozoa is reviewed here and compared against the existing commercial filter options. Because of the differences in sampling procedures, no one method for concentrating these protozoa consistently outperforms others. This comparative article summarises our current understanding of the recent water sampling techniques to concentrate waterborne protozoans and highlights the remaining knowledge gaps. While ultrafiltration is a financially practical alternative for the concentration of pathogens, this paper highlights that further investigation is required to optimise their efficacy, particularly for low numbers of oocysts and cysts. These insights play a substantial role in providing information for advancing enhanced water sampling and treatment systems, reducing the dangers connected with waterborne protozoans, and ensuring the protection of public health. [ABSTRACT FROM AUTHOR]

Giardia duodenalis, an important zoonotic protozoan parasite, adheres to host intestinal epithelial cells (IECs) via the ventral disc and causes giardiasis characterized mainly by diarrhea. To date, it remains elusive how excretory-secretory products (ESPs) of Giardia enter IECs and how the cells respond to the entry. Herein, we initially demonstrated that ESPs evoked IEC endocytosis in vitro. We indicated that ESPs contributed vitally in triggering intrinsic apoptosis, pro-inflammatory responses, tight junction (TJ) protein expressional changes, and autophagy in IECs. Endocytosis was further proven to be implicated in those ESPs-triggered IEC responses. Ten predicted virulent excretory-secretory proteins of G. duodenalis were investigated for their capability to activate clathrin/caveolin-mediated endocytosis (CME/CavME) in IECs. Pyridoxamine 5'-phosphate oxidase (PNPO) was confirmed to be an important contributor. PNPO was subsequently verified as a vital promoter in the induction of giardiasis-related IEC apoptosis, inflammation, and TJ protein downregulation. Most importantly, this process seemed to be involved majorly in PNPO-evoked CME pathway, rather than CavME. Collectively, this study identified Giardia ESPs, notably PNPO, as potentially important pathogenic factors during noninvasive infection. It was also noteworthy that ESPs-evoked endocytosis might play a role in triggering giardiasis-inducing cellular regulation. These findings would deepen our understanding about the role of ESPs, notably PNPO, in the pathogenesis of giardiasis and the potential attributed endocytosis mechanism. [ABSTRACT FROM AUTHOR]

ABSTRACT Giardia duodenalis is a protozoan parasite that infects approximately one billion people worldwide. In this study, the effects of lycopene on experimental giardiasis in mice were investigated by evaluating cyst shedding rate, weight changes, duodenal antioxidant status, and histopathological alteration. Ninety‐five male mice aged four to six weeks were divided into six groups of 15 and one group of 5 as the negative control. All mice were infected with 2 × 105 Giardia duodenalis (B genotype) cysts except for the negative control. The treatment groups were treated with metronidazole, mixed lycopene‐metronidazole, and 5, 10, and 20 mg/kg lycopene for 7 days. The mice treated with lycopene 20 and mixed lycopene‐metronidazole had no cyst shedding from the 7th day of treatment. The mildest lesions were observed in the mixed lycopene‐metronidazole group. Results showed that treatment with mixed lycopene‐metronidazole significantly increased TAC and decreased MDA levels as compared with the positive control. It seems that the antigiardial effect of lycopene is related to its antioxidant properties. Nevertheless, direct effect of lycopene on Giardia and its probable mechanism remain to be investigated. [ABSTRACT FROM AUTHOR]

Dogs offer multiple benefits in their relationship with humans, but they can also be carriers of zoonotic parasites that affect human and animal health. Zoonoses account for about 58% of all human infectious diseases. The objective of this study was to assess intestinal parasitism in dogs with owners in the City of Barranquilla in the years 2016 to 2018. A retrospective descriptive study was carried out that included 3,279 reports of parasitological evaluation of feces from a clinical laboratory that serves a network of veterinary services in the city of Barranquilla. 49.2 % of the dogs had some type of intestinal parasite. The most frequent were helminths: Strongyloides sp. 9.6 %, Toxocara canis 7.7 % and Ancylostoma caninum 6.2 %; and the protozoa: Entamoeba spp. 10.0 %, Isospora spp. 6.9 % and Giardia spp. 5.7 %. The Principal Component Analysis of the parasite profiles by year showed significant differences. The presence of zoonotically transmitted intestinal parasites in dogs evidenced the need to establish corrective and preventive measures in the field of public health that allow their control, since they constitute a significant risk of disease in the community. [ABSTRACT FROM AUTHOR]

Background: Opportunistic microorganisms of the intestinal tract, such as Cryptosporidium spp. Giardia spp. Blastocystis sp. and microsporidia, are increasingly responsible for clinical disorders in various host species, including humans. Objectives: This study was conducted to search for the above parasites in the feces of pet birds using parasitological and molecular methods in Tehran City, Iran. Methods: In the current study, fecal samples of avian birds were collected and investigated with modified Ziehl-Neelsen, modified trichrome, and trichrome staining for the presence of microsporidia, Cryptosporidium, Blastocystis and Giardia. All the samples were examined molecularly with specific primers and PCR methods. Results: Three of the examined droppings contained Encephalitozoon hellem genotype 1B (2%) by PCR and sequencing. The microsporidian organisms were recovered from the droppings of a clinically normal green-cheek parakeet, an African gray parrot, and a lovebird. Other parasites that were examined were not found in the analyzed samples. Conclusion: The current study proved that captive pet birds are a possible source of microsporidian infection. Besides the fact that encephalitozoonosis is predominantly subclinical in birds, the highly resistant nature of the microsporidia spores can put the owners at increased risk of disease acquisition via spore inhalation or ingestion. [ABSTRACT FROM AUTHOR]

Twenty-four treated municipal wastewater samples were collected during one-year study period and examined for the presence of helminthes eggs, Cryptosporidium oocysts, and Giardia cysts to compare between their occurrences. Modified Bailenger method was used in this study to detect and enumerate nematode eggs in the treated wastewater samples, while polymerase chain reaction (PCR) technique was used to detect Cryptosporidium and Giardia in the same treated wastewater samples. Results revealed that Ascaris eggs were detected in only 8.33% out of 24 treated wastewater samples, while Cryptosporidium oocysts and Giardia cysts were detected in 20.83 and 12.5% out of the same 24 treated wastewater samples, respectively. The public's health is at risk since treated wastewater used for agriculture still contains Giardia cysts and Cryptosporidium oocysts, even if it satisfies the nematode criteria. [ABSTRACT FROM AUTHOR]

Endocytosis is a quintessential feature of eukaryotes, and the emergence of endocytosis played a major role in the origin and evolution of eukaryotes. During the early evolution of eukaryotes, phagocytosis and the digestion of prey (i.e., bacteria) combined with the endocytosis of macromolecules opened a new source of nutrients beyond osmotrophy. Pathogenic and commensal protozoa have retained endocytosis as a major mechanism of nutrient acquisition even though, in theory, nutrients could be obtained from the host through osmotrophy. Nearly all pathogenic protozoa exhibit endocytosis and have lysosomal-like compartments that function as digestive vacuoles, and endocytosis appears to play a major role in the acquisition of nutrients. Cryptosporidium is a possible exception that may not exhibit endocytosis. Phagotrophy, however, is only observed in parasites of the intestinal lumen and appears to have been lost in blood and tissue parasites. Overall, the basic features of endocytosis and lysosomes are similar to other eukaryotes. Nonetheless, adaptation to the host has generated some novel features that are specific to certain protozoan lineages. [ABSTRACT FROM AUTHOR]

Toxoplasmosis has a major public health significance because of its social and economic influence on the public. A protozoan parasite causes gastrointestinal problems characterised by acute or chronic diarrhoea, commonly known as giardiasis. The occurrence of giardiasis in dogs and cats has consequences for clinical and public health. Instead of being a potential threat to public health, no study has been planned to detect the occurrence of these important zoonotic parasites in pets of Faisalabad or Pakistan. Therefore, keeping in mind the importance, the current study was planned with the objectives of finding the occurrence of Toxoplasma gondii and Giardia in owned dogs and cats of Faisalabad and to find information about how much the owners know and are concerned about zoonosis. For this purpose, 150 (each for dogs and cats) faecal and blood samples were collected from 150 dogs and cats from pets that were presented to different veterinary clinics. Direct microscopy was used to screen for Giardia, while Toxoplasma was detected using the Toxoplasmosis Latex Kit (ANTEC Diagnostic®, UK). A questionnaire was completed to reveal the associated risk factors and awareness of zoonosis. The generated data were analysed using the chi-squared test. In dogs, an overall prevalence of 42% for Giardia and 37% for Toxoplasma gondii was found. The corresponding values for cats were 36% and 39.33%, respectively. These findings indicated a statistically non-significant (P>0.05) difference in the prevalence of these parasites in both species. Moreover, the findings of the study clearly illustrate that cats and dogs being kept as pet animals pose a serious threat to human beings, and owners are not aware of this alarming situation. [ABSTRACT FROM AUTHOR]

Simple Summary: Non-human primates, due to their close taxonomic relationship with humans, host the highest diversity of parasites with zoonotic potential. In zoological gardens, the presence of helminths and protist parasites with a direct lifecycle is virtually unavoidable; biosecurity measures are of the utmost importance to control their spread and environmental load and avoid transmission to staff and visitors. In this study, we investigated the population of non-human primates in two zoos in Northern Italy to evaluate gastrointestinal parasite diversity and their zoonotic potential. The highest prevalence was registered for protist taxa, i.e., Giardia and Blastocystis. Proof for the transmission of parasites from synanthropic rats to the animals in the collection was also provided. Non-human primates (NHPs) host a variety of helminth and protist parasites that are able to cause infection in humans. Gastrointestinal parasites in NHPs living in two zoological gardens of Northern Italy were studied. An total of 96 faecal pools were collected from 26 groups of NHPs. The mini-Flotac method was applied to fecal samples to detect gastrointestinal helminthiases, while the detection of the protists Cryptosporidium spp., Blastocystis sp. and Giardia duodenalis was performed by targeting SSU rRNA through nested PCR and real-time PCR; they were further studied by sequencing the same gene for Blastocystis and βgiardine and triosephosphate isomerase (TPI) genes for Giardia. Twenty-two out of the 96 examined fecal pools (22.9%) were positive for one or more helminth species, including Hymenolepis diminuta, Trichurid, Capillariid and Strongylid eggs. All samples were negative for Cryptosporidium spp., while 16/26 (61.5%) animals were positive for G. duodenalis in the real-time PCR; the sequences obtained assigned them all to sub-assemblage BIV. Blastocystis sp. was detected in 22/26 of the NHPs (84.6%); molecular analyses attributed the isolates to ST 4, allele 92. Analyses of the feces of sympatric rats revealed the presence of the same allele, as well as of Hymenolepis diminuta eggs, raising concern about their role as parasite reservoirs in the facilities. [ABSTRACT FROM AUTHOR]

Simple Summary: An extensive countrywide investigation was carried out in Greece to study gastrointestinal zoonotic pathogens in horses. Faecal samples were collected and tested. Gastrointestinal pathogens were detected in the faecal samples obtained from 43% of horses examined. They were detected more frequently in samples from horses in courtyard housing. The most frequently detected zoonotic pathogens were enteropathogenic Escherichia coli and Shiga-like toxin-producing E. coli stx1/stx2. The decreasing age of horses and the presence of livestock at the same premises as the horses emerged to be significantly associated with the detection of pathogens. The findings have indicated the presence of pathogens with zoonotic potential in horses and have suggested factors that may play some role in this. The objectives of this study were (a) to detect zoonotic gastrointestinal pathogens in faecal samples of horses using the FilmArray® GI Panel and (b) to identify variables potentially associated with their presence. Faecal samples collected from 224 horses obtained during a countrywide study in Greece were tested by means of the BioFire® FilmArray® Gastrointestinal (GI) Panel, which uses multiplex-PCR technology for the detection of 22 pathogens. Gastrointestinal pathogens were detected in the faecal samples obtained from 97 horses (43.3%). Zoonotic pathogens were detected more frequently in samples from horses in courtyard housing (56.0%) than in samples from horses in other housing types (39.7%) (p = 0.040). The most frequently detected zoonotic pathogens were enteropathogenic Escherichia coli (19.2% of horses) and Shiga-like toxin-producing E. coli stx1/stx2 (13.8%). During multivariable analysis, two variables emerged as significant predictors for the outcome 'detection of at least one zoonotic pathogen in the faecal sample from an animal': (a) the decreasing age of horses (p = 0.0001) and (b) the presence of livestock at the same premises as the horses (p = 0.013). As a significant predictor for the outcome 'detection of two zoonotic pathogens concurrently in the faecal sample from an animal', only the season of sampling of animals (autumn) emerged as significant in the multivariable analysis (p = 0.049). The results indicated a diversity of gastrointestinal pathogens with zoonotic potential in horses and provided evidence for predictors for the infections; also, they can serve to inform horse owners and handlers regarding the possible risk of transmission of pathogens with zoonotic potential. In addition, our findings highlight the importance of continuous surveillance for zoonotic pathogens in domestic animals. [ABSTRACT FROM AUTHOR]

Autoimmune pancreatitis type 2 is a relatively novel entity with some still controversial issues. The current diagnostic algorithm relies on imaging studies and histology. Therapy includes corticosteroids with consequently low risk of relapse in the following year. However, the pathogenesis remains unclear, and data are insufficient for long-term prognosis. We have treated a 17-year-old boy whose autoimmune pancreatitis type 2 was revealed during surgery for a pre-existing biliary tract anomaly with concurrent protozoal infection. We discuss the co-occurrence of these conditions in terms of eventual pathogenesis correlation and combined effect on long-term prognosis. [ABSTRACT FROM AUTHOR]

Cryptosporidium and Giardia are recognized as significant etiological agents of diarrheal outbreaks in humans as these parasites may be transmitted through the ingestion of water and food contaminated with feces of human or animal origin. However, surveillance studies on the role of animal reservoirs in the transmission of Cryptosporidium and Giardia are deemed insufficient and the complete dimension of the problem contributing to contamination in an agricultural setting is unknown. This study aimed to assess the presence of Cryptosporidium and Giardia in domesticated animals from selected farms in the agricultural provinces of Laguna and Quezon in Southern Luzon, Philippines. Using immunofluorescence assay, an overall 85.7% incidence of protozoan infection was recorded among the animals (N = 161). Of these, 77.0 and 73.9% were positive for Cryptosporidium and Giardia, respectively. Highest incidence (95.83%) of Cryptosporidium was documented in swine and the highest incidence (89.47%) of Giardia was observed in ruminants. Analyses revealed significant differences in the incidence of the protozoan parasites among animals with different containment status, water source, age group, and sex. On the other hand, farm workers' knowledge on parasite transmission was negatively correlated (p = 0.001) to parasite incidence. With the scarcity of data about Cryptosporidium and Giardia in farm-raised animals in the Philippines, the information obtained from this study will be vital for protozoan source tracking and further control interventions against Cryptosporidium and Giardia infections. [ABSTRACT FROM AUTHOR]

Giardia duodenalis is the causative agent of the neglected diarrhoeal disease giardiasis. While often self-limiting, giardiasis is ubiquitous and impacts hundreds of millions of people annually. It is also a common gastro-intestinal disease of domestic pets, wildlife, and livestock animals. However, despite this impact, there is no vaccine for Giardia currently available. In addition, treatment relies on chemotherapies that are associated with increasing failure rates. To identify new treatment options for giardiasis we recently screened the Compounds Australia Scaffold Library for new chemotypes with selective anti-Giardia activity, identifying three compounds with sub-μM activity and promising selectivity. Here we extended these studies by examining the anti-Giardia activity of series CL9569 compounds. This compound series was of interest given the promising activity (IC50 1.2 μM) and selectivity demonstrated by representative compound, SN00798525 (1). Data from this work has identified an additional three thieno [3,2-b]pyrrole 5-carboxamides with anti-Giardia activity, including 2 which displayed potent cytocidal (IC50 ≤ 10 nM) and selective activity against multiple Giardia strains, including representatives from both human-infecting assemblages and metronidazole resistant parasites. Preclinical studies in mice also demonstrated that 2 is well-tolerated, does not impact the normal gut microbiota and can reduce Giardia parasite burden in these animals.

Cryptosporidium spp. and Giardia duodenalis are opportunistic zoonotic protozoan parasites related to diarrhea in humans and many mammals. This study aimed to determine the prevalence and genotypes of Cryptosporidium spp. and Giardia duodenalis in shelter dogs in Batman province. The animal material of the study consisted of 100 dogs of different breeds and sexes. Fresh fecal samples taken from the dogs were examined under the microscope by Kinyoun Acid Fast staining for Cryptosporidium spp. and by the native-Lugol method for Giardia. DNA extraction, nested PCR analysis, and sequence analysis were then performed. As a result of the analyses, all samples were negative for Cryptosporidium spp., while Giardia duodenalis was positive in 2% (2/100) of two female dogs less than one-year-old. Sequence analyses of PCR-positive samples showed that the samples overlapped with assemblage C and D samples. Although these results show that shelter dogs in Batman province do not carry a risk for humans in terms of Cryptosporidium spp. and Giardia duodenalis, it is recommended that repeated faecal examinations should be carried out as much as possible to determine the possible role of these parasites in human transmission. [ABSTRACT FROM AUTHOR]

Endocytosis is a quintessential feature of eukaryotes, and the emergence of endocytosis played a major role in the origin and evolution of eukaryotes. During the early evolution of eukaryotes, phagocytosis and the digestion of prey (i.e., bacteria) combined with the endocytosis of macromolecules opened a new source of nutrients beyond osmotrophy. Pathogenic and commensal protozoa have retained endocytosis as a major mechanism of nutrient acquisition even though, in theory, nutrients could be obtained from the host through osmotrophy. Nearly all pathogenic protozoa exhibit endocytosis and have lysosomal-like compartments that function as digestive vacuoles, and endocytosis appears to play a major role in the acquisition of nutrients. Cryptosporidium is a possible exception that may not exhibit endocytosis. Phagotrophy, however, is only observed in parasites of the intestinal lumen and appears to have been lost in blood and tissue parasites. Overall, the basic features of endocytosis and lysosomes are similar to other eukaryotes. Nonetheless, adaptation to the host has generated some novel features that are specific to certain protozoan lineages.

Abstract Giardia duodenalis, a major cause of waterborne infection, infects a wide range of mammalian hosts and is subdivided into eight genetically well-defined assemblages named A through H. However, fragmented genomes and a lack of comparative analysis within and between the assemblages render unclear the molecular mechanisms controlling host specificity and differential disease outcomes. To address this, we generated a near-complete de novo genome of AI assemblage using the Oxford Nanopore platform by sequencing the Be-2 genome. We generated 148,144 long-reads with quality scores of > 7. The final genome assembly consists of only nine contigs with an N50 of 3,045,186 bp. This assembly agrees closely with the assembly of another strain in the AI assemblage (WB-C6). However, a critical difference is that a region previously placed in the five-prime region of Chr5 belongs to Chr4 of Be-2. We find a high degree of conservation in the ploidy, homozygosity, and the presence of cysteine-rich variant-specific surface proteins (VSPs) within the AI assemblage. Our assembly provides a nearly complete genome of a member of the AI assemblage of G. duodenalis, aiding population genomic studies capable of elucidating Giardia transmission, host range, and pathogenicity.

This review explores our understanding of Cryptosporidium species and Giardia duodenalis distribution in Middle East and North African (MENA) water resources. Results emphasize that Cryptosporidium species (sp.) and G. duodenalis (oo)cysts are present in distinct categories of water in ten MENA countries. Cryptosporidium sp. proportional prevalence in the MENA region was 24.5% (95% CI 16.3-33.8), while G. duodenalis prevalence was 37.7% (95% CI 21.9-55.1). Raw wastewater and surface water were the water categories most significantly impacted. Both parasites were reported in the various types of MENA drinking waters. The most frequent species/genotypes reported were C. hominis, C. parvum, and G. duodenalis assemblage A. Despite the high prevalence of (oo)cysts reported, we should consider the absence of waterborne outbreaks. This indicates significant underestimation and underreporting of both parasites in MENA. Stakeholders should apply water contamination legislation to eradicate Cryptosporidium sp. and G. duodenalis (oo)cysts from water resources/categories. [ABSTRACT FROM AUTHOR]

Cryptosporidium and Giardia are infectious parasitic forms widely distributed in aquatic ecosystems and resistant to disinfection of drinking water. Their presence was investigated in the lower areas of the city's four rivers through a four-stage methodology. Between December 2017 and April 2018, three monitoring campaigns were conducted, with results ranging between not detected to 500 oocysts/L for Cryptosporidium, and between not detected and 300 for Giardia. Cryptosporidium was more abundant, especially in the Machángara River. In the same period, the bacteriological quality of the rivers was also reviewed using Total Streptococci and Fecal Enterococci expressed in colony-forming units (CFU)/100 mL as indicators. The results showed a progressive increase in pollution as the course of the rivers progressed. The sensitivity of bacterial indicators to changes in quality is also observed, which is why their use in specific studies is recommended. It is concluded that untreated domestic wastewater discharges may be the main source of contamination by bacteria and parasites and that there is a relationship between their concentration and the seasonal period. In dry weather, the concentration is higher for both microorganisms. This study fills a gap in knowledge in the region, due to the absence of data on parasitic indicators with great impacts on public health. [ABSTRACT FROM AUTHOR]

Introduction: Giardia duodenalis is one of the most important agents of water- and food-borne parasites. The genetic characteristics of this parasite play a crucial role in its pathogenesis. This study represents the first genotyping of Giardia duodenalis in humans using the MLST technique on housekeeping genes, performed in Khuzestan province. Methods: The genomic DNA was extracted from cysts isolated from 129 human fecal specimens, and the diagnosis of G. duodenalis was verified by amplifying a 350 bp fragment of the SSU-rRNA gene. Nested PCR targeting the tpi and β-giardin genes, as well as semi-nested PCR of the gdh gene, were carried out to amplify the 530 bp, 511 bp, and 432 bp fragments of these genes, respectively, followed by sequencing of the amplified fragments. Phylogenetic analysis was then performed to determine the assemblages and sub-assemblages present. Results: Among the 29 specimens that were successfully genotyped for one of the three genes, assemblages A and B were identified in 23 (79.31%) and 6 (20.69%) specimens, respectively. The present study found no significant correlations between assemblages and clinical symptoms, nor with the place of residence. Phylogenetic analysis indicated the presence of subassemblages AII and BIV. Conclusion: The results showed that assemblage A and sub-assemblage AII are the dominant genotypes in Khuzestan province. The identification of sub-assemblages AII and BIV suggests an anthroponotic transmission route. However, further study is necessary to determine the transmission route of Giardia infection from animals to humans. [ABSTRACT FROM AUTHOR]

Cryptosporidium and Giardia are protozoan parasites responsible for gastrointestinal illnesses in humans and in animal species. The main way these parasites are transmitted is by ingestion of their (oo)cysts in drinking water. Monitoring (oo)cysts in water sources is beneficial to evaluate the quality of raw water supplying treatment plants. Currently, the only standardized protocol to enumerate these parasites from water samples is United States Environmental Protection Agency (USEPA) Method 1623.1. With this method, we monitored three major water sources in Quebec over a year to assess temporal and geographical variations of these parasite (oo)cysts. These three water sources have independent watersheds despite being in the same region. We found a general pattern for Giardia, with high concentrations of cysts during cold and transition periods, and significantly lower concentrations during the warm period. Cryptosporidium's concentration was more variable throughout the year. Statistical correlations (Pearson's correlation coefficients) were established between the concentration of each parasite and various environmental parameters. The three study sites each showed unique factors correlating with the presence of both protozoa, supporting the idea that each water source must be seen as a unique entity with its own particular characteristics and therefore, must be monitored independently. Although some environmental parameters could be interesting proxies to the parasitic load, no parameter was strongly correlated throughout the whole sampling year and none of the parameters could be used as a single proxy for all three studies sources. [ABSTRACT FROM AUTHOR]

Ferrate (Fe(VI): HFeO4-/FeO42-), a potent oxidant, has been investigated as an alternative chemical disinfectant in water treatment due to its reduced production of disinfection by-products. In this study, we assessed the disinfecting ability of potassium ferrate against a variety of microorganisms, including waterborne pathogens, under varying pH and water temperature conditions. We presented CT values, a metric of ferrate concentrations (C) and contact time (T), to quantify microbial inactivation rates. Among the tested microorganisms, human adenovirus was the least resistant to ferrate, followed by waterborne bacteria such as Escherichia coli and Vibrio cholerae, and finally, the protozoan parasite Giardia duodenalis. We further investigated the impact of two pH values (7 and 8) and two temperatures (5 and 25 °C) on microbial inactivation rates, observing that inactivation rates increased with lower pH and higher temperature. In addition to showcasing ferrate's capacity to effectively inactivate a range of the tested microorganisms, we offer a ferrate CT table to facilitate the comparison of the effectiveness of various disinfection methods. [ABSTRACT FROM AUTHOR]

Simple Summary: Cryptosporidium spp., Giardia duodenalis, and Enterocytozoon bieneusi are among the most common diarrhea-causing pathogens in humans and livestock. This was the first study to systematically and molecularly detect the prevalence of three intestinal protozoa in cattle in Heilongjiang Province, Northeast China. Our findings indicate that a variety of species/genotypes are prevalent in cattle in Heilongjiang Province and that some of them are zoonotic, indicating a risk of zoonotic disease transmission in endemic areas. This study helps to understand the epidemiology of Cryptosporidium spp., Giardia duodenalis, and Enterocytozoon bieneusi in cattle in Heilongjiang Province. Crytosporidium spp., Giardia duodenalis, and Enterocytozoon bieneusi are important diarrheal pathogens with a global distribution that threatens the health of humans and animals. Despite cattle being potential transmission hosts of these protozoans, the associated risks to public health have been neglected. In the present study, a total of 1155 cattle fecal samples were collected from 13 administrative regions of Heilongjiang Province. The prevalence of Cryptosporidium spp., G. duodenalis, and E. bieneusi were 5.5% (64/1155; 95% CI: 4.2–6.9), 3.8% (44/1155; 95% CI: 2.7–4.9), and 6.5% (75/1155; 95% CI: 5.1–7.9), respectively. Among these positive fecal samples, five Cryptosporidium species (C. andersoni, C. bovis, C. ryanae, C. parvum, and C. occultus), two G. duodenalis assemblages (E and A), and eight E. bieneusi genotypes (BEB4, BEB6, BEB8, J, I, CHS7, CHS8, and COS-I) were identified. Phylogenetic analysis showed that all eight genotypes of E. bieneusi identified in the present study belonged to group 2. It is worth noting that some species/genotypes of these intestinal protozoans are zoonotic, suggesting a risk of zoonotic disease transmission in endemic areas. The findings expanded our understanding of the genetic composition and zoonotic potential of Cryptosporidium spp., G. duodenalis, and E. bieneusi in cattle in Heilongjiang Province. [ABSTRACT FROM AUTHOR]

The sole known heme enzyme of the parasitic protist Giardia intestinalis is a flavohemoglobin (gFlHb) that acts as a nitric oxide dioxygenase (NOD) and protects the organism from the free radical nitric oxide. To learn more about the properties of this enzyme, we measured its nitric oxide dioxygenase, NADH oxidase, and cytochrome c reductase activities and compared these to the activities of the E. coli flavohemoglobin (Hmp). The turnover number for the NOD activity of gFlHb (23 s−1) is about two-thirds of that of Hmp (34 s−1) at pH 6.5 and 37 °C. The two enzymes differ in their sensitivity towards molecules that act as heme ligands. For both gFlHb and Hmp, inhibition with miconazole, a large imidazole ligand, is adequately described by simple competitive inhibition, with KI = 10 μM and 0.27 μM for gFlHb and Hmp, respectively. Inhibition plots with the small ligand imidazole were biphasic, which is consistent with previous experiments with carbon monoxide as a probe that show that the active site of flavohemoglobins exists in two conformations. Interestingly, the largest difference is observed with nitrite, which, like imidazole, also shows a biphasic inhibition plot; however, nitrite inhibits gFlHb at sub-millimolar concentrations while Hmp is not significantly affected. NADH oxidase activity measured under aerobic conditions in the absence of nitric oxide for Hmp was more than twice the activity of gFlHb. The addition of 1 mM hydrogen peroxide in these assays stimulated the NADH oxidase activity of gFlHb but not Hmp. Both enzymes had nearly identical cytochrome c reductase activities but the extent of the contribution of indirect reduction by flavohemoglobin-generated superoxide was much lower with gFlHb (4% SOD-inhibited) than with Hmp (17% SOD-inhibited). Although the active sites of the two enzymes share the same highly conserved residues that are important for catalysis, differences in the distal ligand binding site may account for these differences in activity and sensitivity towards NOD inhibitors. The differences observed in the NADH oxidase and cytochrome c reductase assays suggest that gFlHb may have evolved to protect the protist, which lacks both superoxide dismutase and catalase, from the damaging effects of superoxide by minimizing its production and from peroxide by actively reducing it. [ABSTRACT FROM AUTHOR]

Los perros ofrecen múltiples beneficios en su relación con los humanos, pero también pueden ser portadores de parásitos zoonóticos que afectan la salud humana y animal. Las zoonosis representan alrededor del 58 % de todas las enfermedades infecciosas en los humanos. Se evaluó el parasitismo intestinal en perros con dueños en la ciudad de Barranquilla, Colombia, en los años 2016, 2017 y 2018. Se realizó un estudio descriptivo retrospectivo que incluyó 3,279 reportes de evaluación parasitológica de heces de perro. El 49.2 % de los perros presentó algún tipo de parásito intestinal. Los más frecuentes fueron los helmintos: Strongyloides sp. (9.6%), Toxocara canis (7.7 %) y Ancylostoma caninum (6.2 %); y los protozoarios Entamoeba spp. (10.0 %), Isospora spp. (6.9 %) y Giardia spp. (5.7 %). Un análisis de componentes principales de los perfiles parasitarios por año identificó diferencias significativas entre ellos. La presencia de parásitos intestinales de transmisión zoonótica en los perros domésticos analizados resalta la necesidad de establecer medidas correctivas y preventivas en el ámbito de la salud pública que permitan su control, ya que constituyen un riesgo significativo de enfermedad para los humanas y los perros.

Giardia duodenalis (syn. G. intestinalis, G. lamblia) is a widespread gastrointestinal protozoan parasite with debated taxonomic status. Currently, eight distinct genetic sub-groups, termed assemblages A-H, are defined based on a few genetic markers. Assemblages A and B may represent distinct species and are both of human public health relevance. Genomic studies are scarce and the few reference genomes available, in particular for assemblage B, are insufficient for adequate comparative genomics. Here, by combining long- and short-read sequences generated by PacBio and Illumina sequencing technologies, we provide nine annotated genome sequences for reference from new clinical isolates (four assemblage A and five assemblage B parasite isolates). Isolates chosen represent the currently accepted classification of sub-assemblages AI, AII, BIII and BIV. Synteny over the whole genome was generally high, but we report chromosome-level translocations as a feature that distinguishes assemblage A from B parasites. Orthologue gene group analysis was used to define gene content differences between assemblage A and B and to contribute a gene-set-based operational definition of respective taxonomic units. Giardia is tetraploid, and high allelic sequence heterogeneity (ASH) for assemblage B vs. assemblage A has been observed so far. Noteworthy, here we report an extremely low ASH (0.002%) for one of the assemblage B isolates (a value even lower than the reference assemblage A isolate WB-C6). This challenges the view of low ASH being a notable feature that distinguishes assemblage A from B parasites, and low ASH allowed assembly of the most contiguous assemblage B genome currently available for reference. In conclusion, the description of nine highly contiguous genome assemblies of new isolates of G. duodenalis assemblage A and B adds to our understanding of the genomics and species population structure of this widespread zoonotic parasite.

Undernutrition in children commonly disrupts the structure and function of the small intestinal microbial community, leading to enteropathies, compromised metabolic health, and impaired growth and development. The mechanisms by which diet and microbes mediate the balance between commensal and pathogenic intestinal flora remain elusive. In a murine model of undernutrition, we investigated the direct interactions Giardia lamblia, a prevalent small intestinal pathogen, on indigenous microbiota and specifically on Lactobacillus strains known for their mucosal and growth homeostatic properties. Our research reveals that Giardia colonization shifts the balance of lactic acid bacteria, causing a relative decrease in Lactobacillus spp. and an increase in Bifidobacterium spp. This alteration corresponds with a decrease in multiple indicators of mucosal and nutritional homeostasis. Additionally, protein-deficient conditions coupled with Giardia infection exacerbate the rise of primary bile acids and susceptibility to bile acid-induced intestinal barrier damage. In epithelial cell monolayers, Lactobacillus spp. mitigated bile acid-induced permeability, showing strain-dependent protective effects. In vivo, L. plantarum, either alone or within a Lactobacillus spp consortium, facilitated growth in protein-deficient mice, an effect attenuated by Giardia, despite not inhibiting Lactobacillus colonization. These results highlight Giardia’s potential role as a disruptor of probiotic functional activity, underscoring the imperative for further research into the complex interactions between parasites and bacteria under conditions of nutritional deficiency.

Infection with the protozoan parasite Giardia duodenalis (syn. intestinalis, lamblia) has been associated with intestinal mucus disruptions and microbiota dysbiosis. The mechanisms remain incompletely understood. Mucus consists primarily of densely glycosylated mucin glycoproteins. Mucin O-glycans influence mucus barrier properties and mucin–microbe interactions and are frequently altered during disease. In this study, we observed time-dependent and regiospecific alterations to intestinal mucin glycosylation patterns and the expression of mucin-associated glycosyltransferase genes during Giardia infection. Glycosylation alterations were observed in Giardia-infected mice in the upper small intestine, the site of parasite colonization, and in the distal colon, where active trophozoites were absent. Alterations occurred as early as day 2 post-infection and persisted in mice after parasite clearance. We also observed small intestinal goblet cell hyperplasia and thinning of the distal colon mucus barrier during early infection, and microbiota alterations and altered production of cecal SCFAs. Giardia-induced alterations to mucin glycosylation were at least in part dependent on microbiota dysbiosis, as transplantation of a dysbiotic mucosal microbiota collected from Giardia-infected mice recapitulated some alterations. This study describes a novel mechanism by which Giardia alters intestinal mucin glycosylation, and implicates the small intestinal microbiota in regulation of mucin glycosylation patterns throughout the gastrointestinal tract.

Giardia is a genus of flagellated protozoans that parasitize the gastrointestinal tract of humans and wildlife worldwide. While G. duodenalis is well-studied due to its potential to cause outbreaks of diarrheal illness in humans, other Giardia species from wildlife have been largely understudied. This study examines the occurrence, host specificity, and genotypic diversity of Giardia in wild rodents living within the New York City water supply watershed. A novel nested PCR assay targeting the 18S ssu-rDNA gene is introduced, which captures nearly the entire gene for improved species-level determination versus existing molecular typing methods. Molecular characterization of 55 Giardia specimens reveals at least seven novel lineages. Phylogenetic analysis indicates a close relationship between the newly characterized Giardia lineages and rodent hosts, suggesting rodents as important reservoirs of Giardia and its close relatives. These findings provide insights into the diversity of Giardia species and their public health potential in localities with human-wildlife interaction and further emphasizes the need for continued efforts to improve the molecular tools used to study microbial eukaryotes, especially those with zoonotic potential.

Background Prolonged diarrhoea is common amongst returning travellers and is often caused by intestinal protozoa. However, the epidemiology of travel-associated illness caused by protozoal pathogens is not well described. Methods We analysed records of returning international travellers with illness caused by Giardia duodenalis , Cryptosporidium spp. Cyclospora cayetanensis or Cystoisospora belli , reported to the GeoSentinel Network during January 2007–December 2019. We excluded records of travellers migrating, with an unascertainable exposure country, or from GeoSentinel sites that were not located in high-income countries. Results There were 2517 cases, 82.3% giardiasis (n  = 2072), 11.4% cryptosporidiosis (n  = 287), 6.0% cyclosporiasis (n  = 150) and 0.3% cystoisosporiasis (n  = 8). Overall, most travellers were tourists (64.4%) on long trips (median durations: 18–30 days). Cryptosporidiosis more frequently affected people < 18 years (13.9%) and cyclosporiasis affected people ≥ 40 years (59.4%). Giardiasis was most frequently acquired in South Central Asia (45.8%) and sub-Saharan Africa (22.6%), cryptosporidiosis in sub-Saharan Africa (24.7%) and South-Central Asia (19.5%), cyclosporiasis in South East Asia (31.3%) and Central America (27.3%), and cystoisosporiasis in sub-Saharan Africa (62.5%). Cyclosporiasis cases were reported from countries of uncertain endemicity (e.g. Cambodia) or in countries with no previous evidence of this parasite (e.g. French Guiana). The time from symptom onset to presentation at a GeoSentinel site was the longest amongst travellers with giardiasis (median: 30 days). Over 14% of travellers with cryptosporidiosis were hospitalized. Conclusions This analysis provides new insights into the epidemiology and clinical significance of four intestinal protozoa that can cause morbidity in international travellers. These data might help optimize pretravel advice and post-travel management of patients with travel-associated prolonged gastrointestinal illnesses. This analysis reinforces the importance of international travel-related surveillance to identify sentinel cases and areas where protozoal infections might be undetected or underreported. [ABSTRACT FROM AUTHOR]

Introduction: Iron deficiency anaemia (IDA), the most prevalent type of anaemia, is recognised as a significant global health concern that affects individuals of all ages. Case Presentation: Herein, we present a case involving an adult male coinfected with Helicobacter pylori and Giardia duodenalis, which precipitated severe IDA. Results: A 24-year-old male presented with symptoms including fatigue, dizziness, headache, abdominal pain, and diarrhoea persisting for four weeks. Thorough blood tests, including complete blood counts, blood film, and iron studies, conclusively established the presence of severe IDA. Furthermore, his faecal sample was collected and subjected to analysis of common bacterial and parasitic gastrointestinal infections. Examination of upper and lower gastrointestinal pathogens indicated that the severe IDA was most likely a result of coinfection with H. pylori and G. duodenalis. The patient received treatment involving antibiotics and iron replacement therapy, which resulted in an improvement in both his symptoms and laboratory results. Conclusions: The present report provides crucial insights into the synergistic effect of concurrent H. pylori and G. duodenalis infections, highlighting their potential to induce severe IDA in infected patients. [ABSTRACT FROM AUTHOR]

In this study, the Quantitative Microbial Risk Assessment (QMRA) methodology was applied to estimate the annual risk of Giardia and Cryptosporidium infection associated with a water treatment plant in southern Brazil. The efficiency of the treatment plant in removing protozoa and the effectiveness of the Brazilian legislation on microbiological protection were evaluated, emphasizing the relevance of implementing the QMRA in this context. Two distinct approaches were employed to estimate the mechanical removal of protozoa: The definitions provided by the United States Environmental Protection Agency (USEPA), and the model proposed by Neminski and Ongerth. Although the raw water collected had a higher concentration of Giardia cysts than Cryptosporidium oocysts, the estimated values for the annual risk of infection were significantly higher for Cryptosporidium than for Giardia. From a general perspective, the risk values of protozoa infection were either below or very near the limit set by the World Health Organization (WHO). In contrast, all the risk values of Cryptosporidium infection exceeded the threshold established by the USEPA. Ultimately, it was concluded that the implementation of the QMRA methodology should be considered by the Brazilian authorities, as the requirements and guidelines provided by the Brazilian legislation proved to be insufficient to guarantee the microbiological safety of drinking water. In this context, the QMRA application can effectively contribute to the prevention and investigation of outbreaks of waterborne disease. [ABSTRACT FROM AUTHOR]

This study demonstrates the capability of Raman microscopy for detecting structural differences in Giardia cells exposed to different drugs and incubation times. While metronidazole (MTZ) visibly affects the cells by inducing extracellular vesicle releases of toxic iron intermediates and modified triple-bond moieties, oseltamivir (OSM) alters the phenylalanine and lipid structures. Modifications in the heme protein environment and the transformation of iron from ferric to ferrous observed for both drug treatments are more notable for MTZ. Different contents and amounts of vesicle excretion are detected for 24 h or 48 h with MTZ incubation. At a shorter drug exposure, releases of altered proteins, glycogen, and phospholipids dominate. Agglomerates of transformed iron complexes from heme proteins and multiple-bond moieties prevail at 48 h of treatment. No such vesicle releases are present in the case of OSM usage. Drug incorporations into the cells and their impact on the plasma membrane and the dynamics of lipid raft confirmed by confocal fluorescence microscopy reveal a more destructive extent by OSM, corroborating the Raman results. Raman microscopy provides a broader understanding of the multifaceted factors and mechanisms responsible for giardiasis treatment or drug resistance by enabling a label-free, simultaneous monitoring of structural changes at the cellular and molecular levels. [ABSTRACT FROM AUTHOR]

Background: Cryptosporidium and Giardia are well-known important intestinal zoonotic pathogens that can infect various hosts and cause diarrhoeal diseases. We aimed to determine the epidemiological prevalence and molecular characterization of Cryptosporidium and Giardia species in Himalayan marmot (Marmota himalayana, class Marmota) in the Qinghai Tibetan Plateau Area of Qinghai Province, Northwest China. Methods: Overall, 243 Himalayan marmot fecal samples were collected in 2017 and in 2019 and a two-step nested PCR technique was performed to amplify the fragments of the SSU rRNA gene of Cryptosporidium and 18S ribosomal RNA gene of Giardia. Molecular characterization of Cryptosporidium was performed with the primary primers NDIAGF2 and N-DIAGR2, the secondary primers CPB-DIAGF and CPB-DIAGR. Similarly, molecular characterization of Giardia was used the first primers Gia2029 and Gia2150c, the secondary primers RH11 and RH4. The positive PCR products were sequenced and the sequences were processed by Clustal Omega and BLAST. Phylogenetic analysis was achieved by NJ method in MEGA. Results: The infection rate of Cryptosporidium spp. and G. duodenalis was 4.9% (12/243) and 0.8% (2/243) in M. himalayana, respectively. Cryptosporidium spp. are characterized as novel genotypes Cryptosporidium marmot genotype I (n=3) and Cryptosporidium marmot genotype II (n=9); G. duodenalis assemblage A (n=2) was found in M. himalayana. Conclusion: This is the first report of Cryptosporidium spp. and G. duodenalis infections in M. himalayana in Qinghai of Northwest China. The results indicate the existence of Cryptosporidium species and G. duodenalis infections that may have a potential public health significance. [ABSTRACT FROM AUTHOR]

The objective of this study was to determine the prevalence and to identify the risk factors of Eimeria spp. and Giardia spp. infection in rabbits from the local population of four localities in northern Algeria. Dung samples were collected from 16 farms, totalling 111 rabbits, and were analysed by the flotation method. Additional, data regarding the farms and management practices were obtained by means of a questionnaire used in the surveys and interviews. The results revealed that the prevalence of Eimeriaspp. contamination was 68.75% (11/16) for farms and 58.56% (65/111) for rabbits, respectively. The prevalence of Giardia spp. was respectively 56.25% (9/16) for farms and 11.7% (13/111) for rabbits. The analyses showed that the prevalence of Eimeria spp. was significantly higher in the farms that did not comply with hygiene and non-conventional feeding and watering. However, the prevalence of Giardia spp. was significant in rabbits kept in poor conditions of rearing. In conclusion, this study showed that the prevalence of these two parasites in rabbits from the local population is relevant and may have important implications for the rabbit industry and public health, especially in rural areas. [ABSTRACT FROM AUTHOR]

Giardia duodenalis and Cryptosporidium spp. are important zoonotic protozoan pathogens that infect the gastro-intestinal tract of numerous vertebrates, including humans, and both parasites are responsible for water- or food-borne outbreaks of disease worldwide. Although, globally, both parasites are highly prevalent, particularly in developing countries, epidemiological data from Romania are scarce, and genotyping has rarely been performed. The aims of the present study were to investigate the occurrence and genetic diversity of G. duodenalis and Cryptosporidium spp. in patients hospitalized in Northwestern Romania in relation to clinical and paraclinical presentation and to identify the relative frequency of non-specific symptoms and potential risk factors. Between June 2022 and January 2024, 426 fecal samples were screened for gastro-intestinal parasites by rapid tests and microscopical examination, further confirmed by PCR and sequencing. Giardia duodenalis was detected and characterized in 12 samples (2.82%), while Cryptosporidium parvum was confirmed in four samples (0.94%). A majority of positive patients were symptomatic and reported nausea and vomiting with a significantly higher frequency compared to negative ones. This study provides new insights into the epidemiological status and clinical implications of gastro-intestinal parasite species and genospecies in Romania that are necessary for an in-depth understanding of the potential zoonotic transmission and improvement of patient care. [ABSTRACT FROM AUTHOR]

Background: Cryptosporidiosis and giardiasis are significant parasitic diseases shared between humans and domestic animals. Due to the close contact between humans and domestic animals in rural areas, it is important to consider the potential transmission of zoonotic parasites from infected domestic animals to humans. This investigation aimed to determine the prevalence and molecular characteristics of Cryptosporidium spp. and Giardia duodenalis in domestic animals and villagers. Methods: A total of 116 fecal samples from villagers and 686 fecal samples from domestic animals in Heilongjiang Province, China, were analyzed for two parasites using nested polymerase chain reaction (PCR) targeting various genetic loci and DNA sequence analysis of the PCR products. Results: By sequence analysis of the SSU rRNA gene, the prevalence of Cryptosporidium in humans was 0.9% (1/116), with one species of C. parvum (n = 1) detected; among domestic animals, the prevalence was 2.6% (18/686), with five species identified: C. suis (n = 7) and C. scrofarum (n = 7) in pigs, C. meleagridis (n = 1) in chickens, C. andersoni (n = 1) in cattle, and C. canis (n = 2) in foxes. C. parvum and C. canis were further subtyped as IIdA19G1 and XXa4 on the basis of gp60 gene. Regarding G. duodenalis, based on the SSU rRNA, bg, gdh, and tpi genes, the prevalence in domestic animals was 5.1% (31/608), with three assemblages identified: A (n = 1) in pigs, D (n = 1) in foxes, and E (n = 27) in geese, cattle, pigs, ducks, and sheep, along with mixed infection of A + E (n = 1) in one pig and B + E (n = 1) in one sheep. No G. duodenalis was detected in humans (0/116). Conclusions: The present results show that no overlap of subtypes between animals and villagers was found in Cryptosporidium spp. and G. duodenalis, indicating a minor role of domestic animals in infecting humans in this population. However, the presence of zoonotic protozoa in domestic animals highlights the need for special attention to high-risk individuals during close contact with domestic animals. [ABSTRACT FROM AUTHOR]

Giardia duodenalis is one of the major causes of diarrhea among children. We performed a systematic review and meta-analysis to assess the prevalence of G. duodenalis and associated risk factors among Asian children. We searched online databases (PubMed, Scopus and Web of Science) and Google Scholar search engine for studies published from 1 January 2000 to 15 March 2022 that measured the prevalence of G. duodenalis among Asian children. Accordingly, the pooled prevalence and 95% CIs were estimated using a random-effects meta-analysis model for the included studies. A total of 182 articles from 22 Asian countries met the inclusion criteria. The pooled prevalence of G. duodenalis infection among Asian children was estimated as 15.1% (95% CI 14.1 to 16%). The highest and lowest pooled prevalence values of G. duodenalis infection were estimated for Tajikistan and China as 26.4% (95% CI 22.9 to 30%) and 0.6% (95% CI 0.001 to 1.02%), respectively. The infection had a higher prevalence in males than in females (OR=1.24; 95% CI 1.16 to 1.31; p<0.001), which was statistically significant. Giardiasis is common among Asian children, hence, a prevention and control scheme of this protozoan in children should be considered by health officials and health policymakers, especially in Asian countries where the prevalence is highest. [ABSTRACT FROM AUTHOR]

Purpose: Investigating the genetic variation in thioredoxin reductase (TrxR) and nitroreductase (NR) genes in both treatment-resistant and -sensitive Giardia duodenalis isolates can provide valuable information in identifying potential markers of resistance to metronidazole. The rapid increase in metronidazole treatment failures suggests the presence of genetic resistance mechanisms. By analyzing these genes, researchers can gain insights into the efficacy of metronidazole against G. duodenalis and potentially develop alternative treatment strategies. In this regard, four G. duodenalis isolates (two clinically sensitive and two clinically resistant to metronidazole) were collected from various hospitals of Shiraz, southwestern Iran. Methods: Parasitological methods including sucrose flotation and microscopy were employed for the primary confirmation of G. duodenalis cysts in stool samples. Microscopy-positive samples were approved by SSU-PCR amplification of the parasite DNA. All four positive G. duodenalis specimens at SSU-PCR were afterward analyzed utilizing designed primers based on important metronidazole metabolism genes including TrxR, NR1, and NR2. Results: Unlike TrxR gene, the results of NR1 and NR2 genes showed that there are non-synonymous variations between sequences of treatment-sensitive and -resistant samples compared to reference sequences. Furthermore, the outcomes of molecular docking revealed that there is an interaction between the protein sequence and spatial shape of treatment-resistant samples and metronidazole in the position of serine amino acid based on the NR1 gene. Conclusion: This issue can be one of the possible factors involved in the resistance of Giardia parasites to metronidazole. To reach more accurate results, a large sample size along with simulation and advanced molecular dynamics investigations are needed. [ABSTRACT FROM AUTHOR]

Cryptosporidium spp., Giardia spp. and Enterocytozoon bieneusi are common zoonotic pathogens in humans and animals. Although rodents are important parts of the ecosystem and common hosts for these pathogens, little is known of the distribution, genetic diversity and zoonotic potential of these pathogens in wild rodents. A total of 442 fecal samples were collected from eleven wild rodent species in three provinces of China, and analyzed for these pathogens by PCR and DNA sequencing. The infection rates of Cryptosporidium spp., Giardia spp. and E. bieneusi were 19.9% (88/442), 19.8% (75/378) and 12.2% (54/442), respectively. Altogether, 23 known Cryptosporidium species/genotypes were identified and their distribution varied among different sampling locations or rodent species. Subtyping of the zoonotic Cryptosporidium species identified two novel subtype families XVe and XVf in C. viatorum, the subtype family XIIh and a novel subtype family XIIj in C. ubiquitum, and the subtype family IId in C. parvum. Three Giardia species were identified, including G. microti (n = 57), G. muris (n = 15) and G. duodenalis (n = 3), with G. duodenalis assemblages A and G identified in brown rats in urban areas of Guangdong. In addition, 13 E. bieneusi genotypes including eight known and five novel ones were identified, belonging to Groups 1, 2, 10, 14 and 15. Within nine genotypes in the zoonotic Group 1, common human-pathogenic genotypes D, Type IV, PigEbITS7 and Peru8 were detected only in brown rats and Lesser rice-field rats in urban areas of Guangdong. Apparent host adaptation and geographical differences were observed among Cryptosporidium spp., Giardia spp. and E. bieneusi genotypes in wild rodents in the present study. Furthermore, the zoonotic Cryptosporidium species and E. bieneusi genotypes commonly found here suggest a high zoonotic potential of these pathogens in wild rodents, especially in brown rats in urban areas. Hygiene and One Health measures should be implemented in urban streets and food stores to reduce the possible direct and indirect transmission of these rodent-related pathogens.

Abstract Background Diarrhoeal diseases are common among children in low- and middle-income countries and are major causes of morbidity and mortality. Cryptosporidium and Giardia are considered to be the main parasitic causes of diarrhoea in children. The aim of the present study was to determine the prevalence and associated factors of Cryptosporidium and Giardia infection in children under five years of age presenting at two health centres (Ndirande and Limbe) in Blantyre, Malawi. Methods This cross-sectional study was performed from February to July 2019 and included 972 children under 5 years of age with diarrhoea. Stool samples were immediately tested after collection at enrolment with a rapid diagnostic test for Cryptosporidium and Giardia infection. Descriptive statistics were used to assess the prevalence of these protozoan parasitic infections, and differences in the basic demographic and anthroponotic variables (between children with diarrhoea and parasite infection, being either Cryptosporidium and Giardia or both versus children with diarrhoea but no RDT confirmed parasite infection) were assessed. Their association with Cryptosporidium and Giardia infection was analysed using simple logistic regressions. Results Of the children recruited, 88 (9.1%) tested positive for Cryptosporidium and 184 (18.9%) for Giardia. Children with only a Giardia infection or a coinfection (of both parasites) were significantly older (mean age 24–26 months) compared to children with only a Cryptosporidium infection (mean age 13 months) or no parasitic infection (mean age 14 months). No significant differences were found with respect to gender, body temperature, stunting or wasting between the different groups of children with moderate to severe diarrhoea. Children attending the Ndirande health centre had almost two times higher odds of testing positive for both infections than those attending Limbe health centre. Conclusion Cryptosporidium and Giardia infections are highly prevalent in children

Autoimmune pancreatitis type 2 is a relatively novel entity with some still controversial issues. The current diagnostic algorithm relies on imaging studies and histology. Therapy includes corticosteroids with consequently low risk of relapse in the following year. However, the pathogenesis remains unclear, and data are insufficient for long-term prognosis. We have treated a 17-year-old boy whose autoimmune pancreatitis type 2 was revealed during surgery for a pre-existing biliary tract anomaly with concurrent protozoal infection. We discuss the co-occurrence of these conditions in terms of eventual pathogenesis correlation and combined effect on long-term prognosis.

Non-human primates (NHPs) host a variety of helminth and protist parasites that are able to cause infection in humans. Gastrointestinal parasites in NHPs living in two zoological gardens of Northern Italy were studied. An total of 96 faecal pools were collected from 26 groups of NHPs. The mini-Flotac method was applied to fecal samples to detect gastrointestinal helminthiases, while the detection of the protists Cryptosporidium spp., Blastocystis sp. and Giardia duodenalis was performed by targeting SSU rRNA through nested PCR and real-time PCR; they were further studied by sequencing the same gene for Blastocystis and βgiardine and triosephosphate isomerase (TPI) genes for Giardia. Twenty-two out of the 96 examined fecal pools (22.9%) were positive for one or more helminth species, including Hymenolepis diminuta, Trichurid, Capillariid and Strongylid eggs. All samples were negative for Cryptosporidium spp., while 16/26 (61.5%) animals were positive for G. duodenalis in the real-time PCR; the sequences obtained assigned them all to sub-assemblage BIV. Blastocystis sp. was detected in 22/26 of the NHPs (84.6%); molecular analyses attributed the isolates to ST 4, allele 92. Analyses of the feces of sympatric rats revealed the presence of the same allele, as well as of Hymenolepis diminuta eggs, raising concern about their role as parasite reservoirs in the facilities.

Giardiasis, caused by the protozoan Giardia intestinalis, affects around 400 million people worldwide, emphasizing the critical need for accurate diagnosis to enhance human health, especially in children. Prolonged giardiasis in childhood can lead to intellectual deficits and other complications. A variety of diagnostic tools, including microscopic, immunological, and molecular methods, are available for detecting G. intestinalis infection. Choosing the most suitable method can be challenging due to the abundance of options. This systematic review assesses the reliability and applicability of these diagnostic modalities. Utilizing the Dimensions and Wordart platforms for data analysis, we focus on relevant literature addressing diagnostic methods for human giardiasis. Microscopic techniques, particularly Ritchie's method, emerge as the primary choice, followed by enzyme-linked immunosorbent assay (ELISA) and polymerase chain reaction (PCR). PCR's limited use is attributed to its high cost and infrastructure challenges in developing nations. In conclusion, our analysis supports microscopic methods as the gold standard for giardiasis diagnosis. However, in cases where symptoms persist despite a negative diagnosis, employing more sensitive diagnostic approaches is advisable. [ABSTRACT FROM AUTHOR]

Giardia duodenalis, an important flagellated noninvasive protozoan parasite, infects the upper small intestine and causes a disease termed giardiasis globally. Few members of the heat shock protein (HSP) family have been shown to function as potential defenders against microbial pathogens, while such information is lacking for Giardia. Here we initially screened and indicated that in vitro Giardia challenge induced a marked early upregulation of HSP70 in intestinal epithelial cells (IECs). As noted previously, apoptotic resistance, nitric oxide (NO)-dependent cytostatic effect and parasite clearance, and epithelial barrier integrity represent effective anti-Giardia host defense mechanisms. We then explored the function of HSP70 in modulating apoptosis, NO release, and tight junction (TJ) protein levels in Giardia-IEC interactions. HSP70 inhibition by quercetin promoted Giardia-induced IEC apoptosis, viability decrease, NO release reduction, and ZO-1 and occludin downregulation, while the agonist celastrol could reverse these Giardia-evoked effects. The results demonstrated that HSP70 played a previously unrecognized and important role in regulating anti-Giardia host defense via attenuating apoptosis, promoting cell survival, and maintaining NO and TJ levels. Owing to the significance of apoptotic resistance among those defense-related factors mentioned earlier, we then elucidated the anti-apoptotic mechanism of HSP70. It was evident that HSP70 could negatively regulate apoptosis in an intrinsic way via direct inhibition of Apaf-1 or ROS-Bax/Bcl-2-Apaf-1 axis, and in an extrinsic way via cIAP2-mediated inhibition of RIP1 activity. Most importantly, it was confirmed that HSP70 exerted its host defense function by downregulating apoptosis via Toll-like receptor 4 (TLR4) activation, upregulating NO release via TLR4/TLR2 activation, and upregulating TJ protein expression via TLR2 activation. HSP70 represented a checkpoint regulator providing the crucial link between specific TLR activation and anti-Giardia host defense responses. Strikingly, independent of the checkpoint role of HSP70, TLR4 activation was proven to downregulate TJ protein expression, and TLR2 activation to accelerate apoptosis. Altogether, this study identified HSP70 as a potentially vital defender against Giardia, and revealed its correlation with specific TLR activation. The clinical importance of HSP70 has been extensively demonstrated, while its role as an effective therapeutic target in human giardiasis remains elusive and thus needs to be further clarified. [ABSTRACT FROM AUTHOR]

Background. Giardia duodenalis causes sporadic or epidemic infections in humans. The parasite comprises assemblages A-H with A and B subdivided further into AI-IV and BI-IV subassemblages. Attempts aimed at linking these genotypes with sources and gastrointestinal manifestations of the infection are largely unexplored in rural communities. Methods. In this cross-sectional study, G. duodenalis infection was genotyped and associated with sources, and gastrointestinal signs and symptoms of the disease among residents of Busia County, a rural setting in western Kenya. Demographic and clinical information were captured using standardized forms. Stool specimens were obtained from the patients and used for genotyping at glutamate dehydrogenase and triose-phosphate isomerase loci using the polymerase chain reaction and restriction fragment length polymorphism. Results. Assemblage B (63.6%) was the most prevalent G. duodenalis infection, while A (20.5%) and mixed A/B (15.9%) were also detected. Among the subassemblages, AI (5.7%), AII (8.0%), AIII (3.4), BIII (30.7%), and BIV (17.0%) were diagnosed including the mixed AII/BIII (15.9%), BIII/BIV (15.9%), AI/AIII (2.3%), and AI/AII (1.1%) infections. Binary logistic regression indicated associations for assemblage A with stomach upset, history of nitroimidazole treatment, and residing in a homestead with cattle and B with age < 18 years, history of eating outdoors, vomiting, steatorrhea, and residing in a homestead with cattle, goats, and poultry (p < 0.05 for all). Among the subassemblages, associations were found for AI with residing in a homestead having cattle and history of nitroimidazole treatment, BIII with residing in a homestead having cattle and poultry, and BIV with steatorrhea (p < 0.05 for all). Altogether, this study illustrates that G. duodenalis assemblage B and subassemblage BIII are the most predominant and are linked to age < 18 years, gastrointestinal manifestations, and living in a homestead with domestic ruminants and poultry. Conclusion. Targeted mass prophylactic treatment of domestic animals and utilization of gastrointestinal presentations, age < 18 years, and a history of nitroimidazole use are useful in the diagnosis and prevention of giardiasis among residents of rural communities. [ABSTRACT FROM AUTHOR]

Background and Aim: Giardiasis is a protozoal disease that is globally prevalent in dogs and cats. The clinical manifestations vary, but asymptomatic cases have also been reported. This study aimed to estimate the prevalence of Giardia duodenalis in domestic dogs and cats, characterize the age susceptibility to this disease, and determine the frequency of cases in which cysts are combined with stool changes. Materials and Methods: After centrifugation in a zinc sulfate solution (specific gravity = 1.32 g/cm3), feces of dogs (2761) and cats (1579) were examined microscopically. The age of the animals and the presence of coprological stool characteristics such as fecal odor, consistency, presence of mucus, and other pathological impurities were taken into account. Results: G. duodenalis infection rates were 18.2% (215/1182) in dogs aged 1-12 months and 3.8% (60/1579) in dogs older than 12 months. The infection rate was 7.8% (48/615) in cats aged 1-12 months and 3.35% (33/994) in cats aged >12 months. The most frequently observed coprological abnormalities in cyst-positive animals were soft and/or mushy stool and pungent odor. In dogs, the frequency of these symptoms was 24.4% (67/275), 27.6% (76/275), and 36.4% (100/275) for soft stools, mushy stools, and pungent fecal odor, and 37.8% (31/82), 25.6% (21/82), and 19.5% (16/82), respectively. No stool changes were found when G. duodenalis cysts were detected in dogs in 24.7% (68/275) of cases and in cats in 24.4% (20/82). Conclusion: G. duodenalis cysts are reported more frequently in domestic cats and dogs under 12 months of age than in dogs under 12 months of age. The presence of Giardia cysts is usually accompanied by a strong odor of feces and changes in their consistency. It can be concluded that it is necessary to conduct periodic surveillance for apparently healthy dogs and cats to rule out G. duodenalis infection. [ABSTRACT FROM AUTHOR]

Background: Diarrhoeal diseases are common among children in low- and middle-income countries and are major causes of morbidity and mortality. Cryptosporidium and Giardia are considered to be the main parasitic causes of diarrhoea in children. The aim of the present study was to determine the prevalence and associated factors of Cryptosporidium and Giardia infection in children under five years of age presenting at two health centres (Ndirande and Limbe) in Blantyre, Malawi. Methods: This cross-sectional study was performed from February to July 2019 and included 972 children under 5 years of age with diarrhoea. Stool samples were immediately tested after collection at enrolment with a rapid diagnostic test for Cryptosporidium and Giardia infection. Descriptive statistics were used to assess the prevalence of these protozoan parasitic infections, and differences in the basic demographic and anthroponotic variables (between children with diarrhoea and parasite infection, being either Cryptosporidium and Giardia or both versus children with diarrhoea but no RDT confirmed parasite infection) were assessed. Their association with Cryptosporidium and Giardia infection was analysed using simple logistic regressions. Results: Of the children recruited, 88 (9.1%) tested positive for Cryptosporidium and 184 (18.9%) for Giardia. Children with only a Giardia infection or a coinfection (of both parasites) were significantly older (mean age 24–26 months) compared to children with only a Cryptosporidium infection (mean age 13 months) or no parasitic infection (mean age 14 months). No significant differences were found with respect to gender, body temperature, stunting or wasting between the different groups of children with moderate to severe diarrhoea. Children attending the Ndirande health centre had almost two times higher odds of testing positive for both infections than those attending Limbe health centre. Conclusion: Cryptosporidium and Giardia infections are highly prevalent in children < 5 years with moderate to severe diarrhoea attending the Limbe and Ndirande health centres in Blantyre, Malawi. [ABSTRACT FROM AUTHOR]