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Abstract Background Trypanosomatid parasites are a group of protozoans that cause devastating diseases that disproportionately affect developing countries. These protozoans have developed several mechanisms for adaptation to survive in the mammalian host, such as extensive expansion of multigene families enrolled in host-parasite interaction, adaptation to invade and modulate host cells, and the presence of aneuploidy and polyploidy. Two mechanisms might result in “complex” isolates, with more than two haplotypes being present in a single sample: multiplicity of infections (MOI) and polyploidy. We have developed and validated a methodology to identify multiclonal infections and polyploidy using whole genome sequencing reads, based on fluctuations in allelic read depth in heterozygous positions, which can be easily implemented in experiments sequencing genomes from one sample to larger population surveys. Results The methodology estimates the complexity index (CI) of an isolate, and compares real samples with simulated clonal infections at individual and populational level, excluding regions with somy and gene copy number variation. It was primarily validated with simulated MOI and known polyploid isolates respectively from Leishmania and Trypanosoma cruzi. Then, the approach was used to assess the complexity of infection using genome wide SNP data from 497 trypanosomatid samples from four clades, L. donovani/L. infantum, L. braziliensis, T. cruzi and T. brucei providing an overview of multiclonal infection and polyploidy in these cultured parasites. We show that our method robustly detects complex infections in samples with at least 25x coverage, 100 heterozygous SNPs and where 5–10% of the reads correspond to the secondary clone. We find that relatively small proportions (≤ 7%) of cultured trypanosomatid isolates are complex. Conclusions The method can accurately identify polyploid isolates, and can identify multiclonal infections in scenarios with sufficient genome read coverage. We pack our method in a single R script that requires only a standard variant call format (VCF) file to run ( https://github.com/jaumlrc/Complex-Infections ). Our analyses indicate that multiclonality and polyploidy do occur in all clades, but not very frequently in cultured trypanosomatids. We caution that our estimates are lower bounds due to the limitations of current laboratory and bioinformatic methods.

Objective: This study aimed to retrospectively evaluate the prevalence of protozoan parasites in stool samples collected from patients presenting with various gastrointestinal complaints to the Medical Parasitology Laboratory of Kafkas University Research and Application Hospital between 2019 and 2022. Methods: Stool samples were initially examined using the native-Lugol method for the detection of protozoan parasites, followed by the formol-ethyl acetate sedimentation method, Giemsa, and carbol fuchsin staining methods. Specific immunochromatographic card tests were used for the diagnosis of Entamoeba histolytica, Cryptosporidium spp., and Giardia intestinalis. Results: Of the 2.267 stool samples examined over the four-year period from January 2019 to December 2022, 7.63% were found to contain one or more protozoan parasites. Among these parasites, Entamoeba histolytica was detected at the highest rate of 4.06%. The other parasite species were identified as follows: Blastocystis spp. 1.15%, Entamoeba spp. and Entamoeba coli each 0.52%, Giardiaintestinalis 0.48%, Endolimax nana 0.17%, and Entamoeba histolytica/dispar 0.08%. Conclusion: This study indicates that despite a decrease in the prevalence of intestinal protozoan infections in the Kars region, these infections remain a significant public health issue. Therefore, improvements in hygiene and sanitation conditions, increased public health education, and the widespread implementation of early diagnosis and treatment methods are necessary. Special measures should be taken to protect vulnerable groups, particularly children and the elderly.

Background: Trypanosomatid parasites are a group of protozoans that cause devastating diseases that disproportionately affect developing countries. These protozoans have developed several mechanisms for adaptation to survive in the mammalian host, such as extensive expansion of multigene families enrolled in host-parasite interaction, adaptation to invade and modulate host cells, and the presence of aneuploidy and polyploidy. Two mechanisms might result in "complex" isolates, with more than two haplotypes being present in a single sample: multiplicity of infections (MOI) and polyploidy. We have developed and validated a methodology to identify multiclonal infections and polyploidy using whole genome sequencing reads, based on fluctuations in allelic read depth in heterozygous positions, which can be easily implemented in experiments sequencing genomes from one sample to larger population surveys. Results: The methodology estimates the complexity index (CI) of an isolate, and compares real samples with simulated clonal infections at individual and populational level, excluding regions with somy and gene copy number variation. It was primarily validated with simulated MOI and known polyploid isolates respectively from Leishmania and Trypanosoma cruzi. Then, the approach was used to assess the complexity of infection using genome wide SNP data from 497 trypanosomatid samples from four clades, L. donovani/L. infantum, L. braziliensis, T. cruzi and T. brucei providing an overview of multiclonal infection and polyploidy in these cultured parasites. We show that our method robustly detects complex infections in samples with at least 25x coverage, 100 heterozygous SNPs and where 5–10% of the reads correspond to the secondary clone. We find that relatively small proportions (≤ 7%) of cultured trypanosomatid isolates are complex. Conclusions: The method can accurately identify polyploid isolates, and can identify multiclonal infections in scenarios with sufficient genome read coverage. We pack our method in a single R script that requires only a standard variant call format (VCF) file to run (https://github.com/jaumlrc/Complex-Infections). Our analyses indicate that multiclonality and polyploidy do occur in all clades, but not very frequently in cultured trypanosomatids. We caution that our estimates are lower bounds due to the limitations of current laboratory and bioinformatic methods. [ABSTRACT FROM AUTHOR]

Peptidyl-prolyl cis/trans isomerases (PPIases) are present in a wide variety of microorganisms, including protozoan parasites such as Trypanosoma cruzi, Trypanosoma brucei, Trichomonas vaginalis, Leishmania major, Leishmania donovani, Plasmodium falciparum, Plasmodium vivax, Entamoeba histolytica, Giardia intestinalis, Cryptosporidium parvum, and Cryptosporidium hominis, all of which cause important neglected diseases. PPIases are classified as cyclophilins, FKBPs, or parvulins and play crucial roles in catalyzing the cis-trans isomerization of the peptide bond preceding a proline residue. This activity assists in correct protein folding. However, experimentally, the biological structure–function characterization of PPIases from these protozoan parasites has been poorly addressed. The recombinant production of these enzymes is highly relevant for this ongoing research. Thus, this review explores the structural diversity, functions, recombinant production, activity, and inhibition of protozoan PPIases. We also highlight their potential as biotechnological tools for the in vitro refolding of other recombinant proteins from these parasites. These applications are invaluable for the development of diagnostic and therapeutic tools. [ABSTRACT FROM AUTHOR]

Abstract Background Enteric parasitic infections remain a major public health problem globally. Cryptosporidium spp., Cyclospora spp. and Giardia spp. are parasites that cause diarrhea in the general populations of both developed and developing countries. Information from molecular genetic studies on the speciation of these parasites and on the role of animals as vectors in disease transmission is lacking in Ghana. This study therefore investigated these diarrhea-causing parasites in humans, domestic rats and wildlife animals in Ghana using molecular tools. Methods Fecal samples were collected from asymptomatic school children aged 9–12 years living around the Shai Hills Resource Reserve (tourist site), from wildlife (zebras, kobs, baboons, ostriches, bush rats and bush bucks) at the same site, from warthogs at the Mole National Park (tourist site) and from rats at the Madina Market (a popular vegetable market in Accra, Ghana. The 18S rRNA gene (18S rRNA) and 60-kDa glycoprotein gene (gp60) for Cryptosporidium spp., the glutamate dehydrogenase gene (gdh) for Giardia spp. and the 18S rDNA for Cyclospora spp. were analyzed in all samples by PCR and Sanger sequencing as markers of speciation and genetic diversity. Results The parasite species identified in the fecal samples collected from humans and animals included the Cryptosporidium species C. hominis, C. muris, C. parvum, C. tyzzeri, C. meleagridis and C. andersoni; the Cyclopora species C. cayetanensis; and the Gardia species, G. lamblia and G. muris. For Cryptosporidium, the presence of the gp60 gene confirmed the finding of C. parvum (41%, 35/85 samples) and C. hominis (29%, 27/85 samples) in animal samples. Cyclospora cayetanensis was found in animal samples for the first time in Ghana. Only one human sample (5%, 1/20) but the majority of animal samples (58%, 51/88) had all three parasite species in the samples tested. Conclusions Based on these results of fecal sample testing for parasites, we conclude that animals and human share species of the three genera (Cryptosporidium, Cyclospora, Giardia), with the parasitic species mostly found in animals also found in human samples, and vice-versa. The presence of enteric parasites as mixed infections in asymptomatic humans and animal species indicates that they are reservoirs of infections. This is the first study to report the presence of C. cayetanensis and C. hominis in animals from Ghana. Our findings highlight the need for a detailed description of these parasites using high-throughput genetic tools to further understand these parasites and the neglected tropical diseases they cause in Ghana where such information is scanty. Graphical Abstract

Entamoeba histolytica is the protozoan causative of human amoebiasis. The EhADH adhesin (687 aa) is a protein involved in tissue invasion, phagocytosis and host-cell lysis. EhADH adheres to the prey and follows its arrival to the multivesicular bodies. It is an accessory protein of the endosomal sorting complexes required for transport (ESCRT) machinery. Here, to study the role of different parts of EhADH during virulence events, we produced trophozoites overexpressing the three domains of EhADH, Bro1 (1–400 aa), Linker (246–446 aa) and Adh (444–687 aa) to evaluate their role in virulence. The TrophozBro11–400 slightly increased adherence and phagocytosis, but these trophozoites showed a higher ability to destroy cell monolayers, augment the permeability of cultured epithelial cells and mouse colon, and produce more damage to hamster livers. The TrophozLinker226–446 also increased the virulence properties, but with lower effect than the TrophozBro11–400. In addition, this fragment participates in cholesterol transport and GTPase binding. Interestingly, the TrophozAdh444–687 produced the highest effect on adherence and phagocytosis, but it poorly influenced the monolayers destruction; nevertheless, they augmented the colon and liver damage. To identify the protein partners of each domain, we used recombinant peptides. Pull-down assays and mass spectrometry showed that Bro1 domain interplays with EhADH, Gal/GalNAc lectin, EhCPs, ESCRT machinery components and cytoskeleton proteins. While EhADH, ubiquitin, EhRabB, EhNPC1 and EhHSP70 were associated to the Linker domain, and EhADH, EhHSP70, EhPrx and metabolic enzymes interacted to the Adh domain. The diverse protein association confirms that EhADH is a versatile molecule with multiple functions probably given by its capacity to form distinct molecular complexes. [ABSTRACT FROM AUTHOR]

Background: Enteric parasitic infections remain a major public health problem globally. Cryptosporidium spp., Cyclospora spp. and Giardia spp. are parasites that cause diarrhea in the general populations of both developed and developing countries. Information from molecular genetic studies on the speciation of these parasites and on the role of animals as vectors in disease transmission is lacking in Ghana. This study therefore investigated these diarrhea-causing parasites in humans, domestic rats and wildlife animals in Ghana using molecular tools. Methods: Fecal samples were collected from asymptomatic school children aged 9–12 years living around the Shai Hills Resource Reserve (tourist site), from wildlife (zebras, kobs, baboons, ostriches, bush rats and bush bucks) at the same site, from warthogs at the Mole National Park (tourist site) and from rats at the Madina Market (a popular vegetable market in Accra, Ghana. The 18S rRNA gene (18S rRNA) and 60-kDa glycoprotein gene (gp60) for Cryptosporidium spp., the glutamate dehydrogenase gene (gdh) for Giardia spp. and the 18S rDNA for Cyclospora spp. were analyzed in all samples by PCR and Sanger sequencing as markers of speciation and genetic diversity. Results: The parasite species identified in the fecal samples collected from humans and animals included the Cryptosporidium species C. hominis, C. muris, C. parvum, C. tyzzeri, C. meleagridis and C. andersoni; the Cyclopora species C. cayetanensis; and the Gardia species, G. lamblia and G. muris. For Cryptosporidium, the presence of the gp60 gene confirmed the finding of C. parvum (41%, 35/85 samples) and C. hominis (29%, 27/85 samples) in animal samples. Cyclospora cayetanensis was found in animal samples for the first time in Ghana. Only one human sample (5%, 1/20) but the majority of animal samples (58%, 51/88) had all three parasite species in the samples tested. Conclusions: Based on these results of fecal sample testing for parasites, we conclude that animals and human share species of the three genera (Cryptosporidium, Cyclospora, Giardia), with the parasitic species mostly found in animals also found in human samples, and vice-versa. The presence of enteric parasites as mixed infections in asymptomatic humans and animal species indicates that they are reservoirs of infections. This is the first study to report the presence of C. cayetanensis and C. hominis in animals from Ghana. Our findings highlight the need for a detailed description of these parasites using high-throughput genetic tools to further understand these parasites and the neglected tropical diseases they cause in Ghana where such information is scanty. [ABSTRACT FROM AUTHOR]

Protozoan parasites cause a wide range of devastating diseases in various kinds of organisms, including humans. It may be lethal if untreated promptly. To detect specific disease-causingorganisms parasites, a wide range of immunological and molecular technologies are now widely available. However, all of this depends on the worker's expertise and are time-consuming, error-prone, and expensive. With the development of technology, compared to traditional biological techniques, convolutional neural networks have reached excellent achievements in image classification, cutting costs while attaining an overall higher accuracy and eliminating human error. Many models include numerous convolutional layers and offer an accuracy between 90 and 95 percent. In this study, 4740 microscopic images of protozoan parasites from six classes with a balanced dataset and an 80–20% split were classified using three convolutional layers with stochastic gradient descent as an optimizer. A 5-fold cross-validation approach is used to evaluate the proposed method. We also examine and evaluate with deep learning models namely VGG16, ResNet50, and InceptionV3. The performance evaluation of the proposed model shows an accuracy of 94% with a precision range (of 0.83-0.99) and a recall range (of 0.76-1.00), respectively. The retrained model was able to recognize and classify all 6 different parasites. Except for class Leishmania, where 24% of images are incorrectly classified as Plasmodium and Trichomonas, the model demonstrates that most cases are correctly identified.

Abstract Background Patients with mental disorders have a high risk of intestinal parasitic infection due to poor hygiene practices. Hence, to better clarify this overlooked phenomenon, the current study is conducted to determine the global prevalence of protozoan parasite infections in patients with mental disorders and investigate the associated risk factors. Methods Several databases (PubMed, Scopus, Web of Science, ProQuest, and Google Scholar) were searched for papers published until December 2022. The fixed effect meta-analysis was used to estimate the overall odds ratio (OR) and pooled prevalence was estimated using a random-effects model with a 95% confidence interval (CI). Results Totally, 131 articles (91 case–control and 40 cross-sectional studies) met the eligibility criteria. Patients with mental disorders were significantly at higher risk for protozoan parasites than healthy controls (OR: 2.059, 1.830–2.317). The highest pooled OR (2.485, 1.413–4.368) was related to patients with neurodevelopmental disorders, and the highest pooled prevalence was detected in patients with neurodevelopmental disorders (0.341, 0.244–0.446), followed by bipolar and related disorders (0.321, 0.000–0.995). Toxoplasma gondii was the most prevalent protozoan parasite (0.343, 0.228–0.467) in cross-sectional studies and the highest pooled OR was related to Cyclospora cayetanensis (4.719, 1.352–16.474) followed by Cryptosporidium parvum (4.618, 2.877–7.412). Conclusion Our findings demonstrated that individuals afflicted with mental disorders are significantly more susceptible to acquiring protozoan parasites in comparison to healthy individuals. Preventive interventions, regular screening, and treatment approaches for parasitic diseases should be considered for patients with mental disorders.

This document is an editorial published in the journal Frontiers in Cellular & Infection Microbiology. The editorial discusses the use of stem cell technologies in the study of tropical infectious diseases. The authors highlight the challenges in studying these diseases and the potential of stem cell technologies to overcome these challenges. They provide examples of how stem cell technologies have been applied to the study of malaria and Chagas disease, and they discuss the potential of these technologies in studying other parasites such as Toxoplasma gondii and helminths. The authors emphasize the importance of stem cell-based approaches in understanding and finding new avenues for prevention and treatment of these diseases. [Extracted from the article]

The current study presents a comprehensive review of worldwide waterborne parasitic protozoan outbreaks reported between 2017 and 2022. In total, 416 outbreaks were attributed to the waterborne transmission of parasitic protozoa. Cryptosporidium accounted for 77.4% (322) of outbreaks, while Giardia was identified as the etiological agent in 17.1% (71). Toxoplasma gondii and Naegleria fowleri were the primary causes in 1.4% (6) and 1% (4) of outbreaks, respectively. Blastocystis hominis, Cyclospora cayetanensis, and Dientamoeba fragilis were independently identified in 0.72% (3) of outbreaks. Moreover, Acanthamoeba spp., Entamoeba histolytica, Vittaforma corneae, and Enterocytozoon bieneusi were independently the causal agents in 0.24% (1) of the total outbreaks. The majority of the outbreaks (195, 47%) were reported in North America. The suspected sources for 313 (75.2%) waterborne parasitic outbreaks were recreational water and/or swimming pools, accounting for 92% of the total Cryptosporidium outbreaks. Furthermore, 25.3% of the outbreaks caused by Giardia were associated with recreational water and/or swimming pools. Developing countries are most likely to be impacted by such outbreaks due to the lack of reliable monitoring strategies and water treatment processes. There is still a need for international surveillance and reporting systems concerning both waterborne diseases and water contamination with parasitic protozoa. HIGHLIGHTS 416 waterborne protozoan outbreaks were identified between 2017 and 2022, globally.; Cryptosporidium was the most common etiological agent.; The majority of the outbreaks were reported in North America.; The number of reported waterborne outbreaks in developed countries were higher than developing countries.; Recreational water and/or swimming pools were the suspected source of infection in most of the reports.;

Parasitic protozoa are single-celled organisms that have adapted to live in cells of humans and animals. The protozoan parasites include Leishmania spp., Trypanosoma spp., Plasmodium spp., Toxoplasma gondii, Cryptosporidium spp., and Eimeria spp., which can cause protozoal diseases that seriously endanger human and animal health and cause huge economic losses in the livestock industry. Parasite development and reproduction after invasion of the host require a large number of purine nucleotides, and the corresponding purine bases are catalyzed by purine phosphoribosyl transferase to generate the corresponding purine nucleotides. Purine phosphoribosyltransferases (PPRT) are important metabolic enzymes involved in the ribophosphorylation of purine bases and are present in many protozoan parasites. Adenine phosphoribosyltransferase (APRT) and hypoxanthine-guanine-xanthine phosphoribosyltransferase (HGXPRT) belong to PPRT. APRT and HGXPRT convert adenine, hypoxanthine, guanine, and xanthine into adenosine-5'-monophosphate (AMP), inosine-5 '-monophosphate (IMP), guanosine-5'-monophosphate (GMP), and xanthosine- 5'-monophosphate (XMP), respectively. Purine nucleotides are involved in many functions as components of DNA and RNA, as enzyme cofactors in metabolic pathways, as sources of energy in protozoan parasites. Purine phosphoribosyltransferases are key enzyme in the purine salvage pathway of parasitic protozoa, which is significantly different from the de novo synthesis pathway of the host. In recent years, purine phosphoribosyltransferases have become the interesting research hotspot as an antiparasitic drug candidates target, with major progress in the screening of compounds against protozoan parasites. This review focuses on the basic characteristics, biological functions, inhibitors screening and application of purine phosphoribosyltransferases in parasitic protozoa of Leishmania spp., Trypanosoma spp., Plasmodium spp., and Toxoplasma gondii, so as to provide reference for the research of drug targets and the screening of new inhibitors against parasitic protozoa.

Background: Patients with mental disorders have a high risk of intestinal parasitic infection due to poor hygiene practices. Hence, to better clarify this overlooked phenomenon, the current study is conducted to determine the global prevalence of protozoan parasite infections in patients with mental disorders and investigate the associated risk factors. Methods: Several databases (PubMed, Scopus, Web of Science, ProQuest, and Google Scholar) were searched for papers published until December 2022. The fixed effect meta-analysis was used to estimate the overall odds ratio (OR) and pooled prevalence was estimated using a random-effects model with a 95% confidence interval (CI). Results: Totally, 131 articles (91 case–control and 40 cross-sectional studies) met the eligibility criteria. Patients with mental disorders were significantly at higher risk for protozoan parasites than healthy controls (OR: 2.059, 1.830–2.317). The highest pooled OR (2.485, 1.413–4.368) was related to patients with neurodevelopmental disorders, and the highest pooled prevalence was detected in patients with neurodevelopmental disorders (0.341, 0.244–0.446), followed by bipolar and related disorders (0.321, 0.000–0.995). Toxoplasma gondii was the most prevalent protozoan parasite (0.343, 0.228–0.467) in cross-sectional studies and the highest pooled OR was related to Cyclospora cayetanensis (4.719, 1.352–16.474) followed by Cryptosporidium parvum (4.618, 2.877–7.412). Conclusion: Our findings demonstrated that individuals afflicted with mental disorders are significantly more susceptible to acquiring protozoan parasites in comparison to healthy individuals. Preventive interventions, regular screening, and treatment approaches for parasitic diseases should be considered for patients with mental disorders. [ABSTRACT FROM AUTHOR]

Stochastic bottlenecks encountered during infection can have a disproportionately large influence on successful parasite colonization. The relationship between parasite infection of discrete tissue niches and long-term persistence or transmission is not fully understood. Cellular barcoding combined with spatial proteomics and transcriptomics has the potential to connect gene expression phenotypes with infection outcomes during colonization. Protozoan pathogens such as Plasmodium spp., Leishmania spp., Toxoplasma gondii , and Trypanosoma spp. are often associated with high-mortality, acute and chronic diseases of global health concern. For transmission and immune evasion, protozoans have evolved diverse strategies to interact with a range of host tissue environments. These interactions are linked to disease pathology, yet our understanding of the association between parasite colonization and host homeostatic disruption is limited. Recently developed techniques for cellular barcoding have the potential to uncover the biology regulating parasite transmission, dissemination, and the stability of infection. Understanding bottlenecks to infection and the in vivo tissue niches that facilitate chronic infection and spread has the potential to reveal new aspects of parasite biology. [ABSTRACT FROM AUTHOR]

Cryptosporidium parvum is a protozoan parasite and one of the leading causes of gastroenteritis in the world, primarily affecting very young children and immunocompromised patients. While infection is usually self-limiting, it can become chronic and even lethal in these vulnerable populations, in whom Cryptosporidium treatments are generally ineffective, due to their acting in concert with a functioning immune system. Here, we describe a case of chronic cryptosporidiosis in a European child with severe CD40L immunodeficiency infected with Cryptosporidium parvum of the IIa20G1 subgenotype, a lineage which has thus far only ever been described in the Middle East. After years of on-off treatment with conventional and non-conventional anti-parasitic drugs failed to clear parasitosis, we performed targeted metagenomics to observe the bacterial composition of the patient's gut microbiota (GM), and to evaluate fecal microbiota transplantation (FMT) as a potential treatment option. We found that C. parvum infection led to significant shifts in GM bacterial composition in our patient, with consequent shifts in predicted intestinal functional signatures consistent with a state of persistent inflammation. This, combined with the patient's poor prognosis and increasing parasitic burden despite many rounds of anti-parasitic drug treatments, made the patient a potential candidate for an experimental FMT procedure. Unfortunately, given the many comorbidities that were precipitated by the patient's immunodeficiency and chronic C. parvum infection, FMT was postponed in favor of more urgently necessary liver and bone marrow transplants. Tragically, after the first liver transplant failed, the patient lost his life before undergoing FMT and a second liver transplant. With this case report, we present the first description of how cryptosporidiosis can shape the gut microbiota of a pediatric patient with severe immunodeficiency. Finally, we discuss how both our results and the current scientific literature suggest that GM modulations, either by probiotics or FMT, can become novel treatment options for chronic Cryptosporidium infection and its consequent complications, especially in those patients who do not respond to the currently available anti-parasitic therapies. [ABSTRACT FROM AUTHOR]

A study on the protozoan parasites and bacterial contamination of surfaces in selected public restrooms in markets, motor parks and students’ hostels in Benin City, southern Nigeria. Out of the 200 swab samples from all the restroom surfaces examined, 15(7.5%) and 176 (88%) were positive for protozoan parasites and bacterial contamination, respectively. There was no significant difference between the female and male restrooms for both (a) protozoan parasites contamination (11% vs 4%) respectively and (b) bacterial contamination (94% vs 82%) respectively. Among the four types of restroom surfaces assessed, toilet seats recorded the highest prevalence, 12.5% of positive samples contamination for protozoan parasites. For bacterial contaminants, the restroom floor had the highest prevalence, 92.5% of positivity as well as the highest variety of bacterial species (eight of the nine species isolated). Although restrooms in markets had the highest positive samples contamination for both protozoan parasites and bacteria, 11% vs 100%, respectively, only bacterial contamination recorded a significant association (P < 0.01). The cysts/oocysts of six species of protozoan parasites contaminants, namely: Entamoeba histolytica, E. coli, Giardia intestinalis, Cryptosporidium sp., Cystisospora belli and Balantidium coli, were recovered during the study. Nine species of bacterial contaminants, namely: Staphylococcus aureus, Escherichia coli, Klebsiella spp., Streptococcus pyogenes, Enterobacter spp., Micrococcus spp., Pseudomonas aeruginosa, Salmonella spp. and Proteus sp., were isolated. Entamaeba histolytica and Staphylococcus aureus recorded the highest prevalence, 33.3% vs 25%, for protozoan parasites and bacterial contaminants respectively. The public health effects associated with the infection of humans by these protozoan and bacterial pathogens are highlighted. The isolation of the pathogens in the study indicated that the routine cleaning and decontamination of the public restrooms were inadequate and the indoor surfaces less than sanitary. Recommendations are proffered on measures to effectively reduce the risk of pathogen transmission or acquisition in public restrooms in the study area. [ABSTRACT FROM AUTHOR]

The article in the Journal of Extracellular Vesicles discusses the global health challenge posed by parasitic infections, affecting over a billion people annually. It highlights the importance of extracellular vesicles (EVs) secreted by parasites in influencing host-parasite interactions and disease progression. The article emphasizes the need for standardized protocols for isolating and characterizing EVs to advance research in parasitology and improve health outcomes worldwide. [Extracted from the article]

Peptidyl-prolyl cis/trans isomerases (PPIases) are present in a wide variety of microorganisms, including protozoan parasites such as Trypanosoma cruzi, Trypanosoma brucei, Trichomonas vaginalis, Leishmania major, Leishmania donovani, Plasmodium falciparum, Plasmodium vivax, Entamoeba histolytica, Giardia intestinalis, Cryptosporidium parvum, and Cryptosporidium hominis, all of which cause important neglected diseases. PPIases are classified as cyclophilins, FKBPs, or parvulins and play crucial roles in catalyzing the cis-trans isomerization of the peptide bond preceding a proline residue. This activity assists in correct protein folding. However, experimentally, the biological structure–function characterization of PPIases from these protozoan parasites has been poorly addressed. The recombinant production of these enzymes is highly relevant for this ongoing research. Thus, this review explores the structural diversity, functions, recombinant production, activity, and inhibition of protozoan PPIases. We also highlight their potential as biotechnological tools for the in vitro refolding of other recombinant proteins from these parasites. These applications are invaluable for the development of diagnostic and therapeutic tools.

The occurrence of Giardia and Cryptosporidium (oo)cysts in drinking source water poses a serious public health risk. Here, we established a method that combines membrane concentration and real-time polymerase chain reaction (PCR) to quantify Giardia and Cryptosporidium in drinking water. The water samples were filtered through a cellulose membrane to collect Giardia and Cryptosporidium, and then nucleic acids were extracted. Specific primers and probes were designed and synthesized according to the gph gene sequence of Giardia and 18S rRNA gene sequence of Cryptosporidium. The concentrations of the two targets were determined using real-time PCR technology. The sensitivity, specificity, and stability of the method were evaluated. Our findings revealed that the detection limits of real-time PCR method for detecting Giardia and Cryptosporidium were 0.926 and 0.65 copy/µL, respectively; the spiked recovery rates were above 60% and 38%, respectively, and relative standard deviations were under 0.95% and 2.26%, respectively. Therefore, this effective procedure based on the membrane concentration method and real-time PCR will be useful for detecting Giardia and Cryptosporidium in drinking water for purpose of continuous environmental monitoring. [ABSTRACT FROM AUTHOR]

Parasites are responsible for the most neglected tropical diseases, affecting over a billion people worldwide (WHO, 2015) and accounting for billions of cases a year and responsible for several millions of deaths. Research on extracellular vesicles (EVs) has increased in recent years and demonstrated that EVs shed by pathogenic parasites interact with host cells playing an important role in the parasite's survival, such as facilitation of infection, immunomodulation, parasite adaptation to the host environment and the transfer of drug resistance factors. Thus, EVs released by parasites mediate parasite‐parasite and parasite‐host intercellular communication. In addition, they are being explored as biomarkers of asymptomatic infections and disease prognosis after drug treatment. However, most current protocols used for the isolation, size determination, quantification and characterization of molecular cargo of EVs lack greater rigor, standardization, and adequate quality controls to certify the enrichment or purity of the ensuing bioproducts. We are now initiating major guidelines based on the evolution of collective knowledge in recent years. The main points covered in this position paper are methods for the isolation and molecular characterization of EVs obtained from parasite‐infected cell cultures, experimental animals, and patients. The guideline also includes a discussion of suggested protocols and functional assays in host cells [ABSTRACT FROM AUTHOR]

The current study presents a comprehensive review of worldwide waterborne parasitic protozoan outbreaks reported between 2017 and 2022. In total, 416 outbreaks were attributed to the waterborne transmission of parasitic protozoa. Cryptosporidium accounted for 77.4% (322) of outbreaks, while Giardia was identified as the etiological agent in 17.1% (71). Toxoplasma gondii and Naegleria fowleri were the primary causes in 1.4% (6) and 1% (4) of outbreaks, respectively. Blastocystis hominis, Cyclospora cayetanensis, and Dientamoeba fragilis were independently identified in 0.72% (3) of outbreaks. Moreover, Acanthamoeba spp., Entamoeba histolytica, Vittaforma corneae, and Enterocytozoon bieneusi were independently the causal agents in 0.24% (1) of the total outbreaks. The majority of the outbreaks (195, 47%) were reported in North America. The suspected sources for 313 (75.2%) waterborne parasitic outbreaks were recreational water and/or swimming pools, accounting for 92% of the total Cryptosporidium outbreaks. Furthermore, 25.3% of the outbreaks caused by Giardia were associated with recreational water and/or swimming pools. Developing countries are most likely to be impacted by such outbreaks due to the lack of reliable monitoring strategies and water treatment processes. There is still a need for international surveillance and reporting systems concerning both waterborne diseases and water contamination with parasitic protozoa. [ABSTRACT FROM AUTHOR]

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Cryptosporidium parvum is a protozoan parasite and one of the leading causes of gastroenteritis in the world, primarily affecting very young children and immunocompromised patients. While infection is usually self-limiting, it can become chronic and even lethal in these vulnerable populations, in whom Cryptosporidium treatments are generally ineffective, due to their acting in concert with a functioning immune system. Here, we describe a case of chronic cryptosporidiosis in a European child with severe CD40L immunodeficiency infected with Cryptosporidium parvum of the IIa20G1 subgenotype, a lineage which has thus far only ever been described in the Middle East. After years of on-off treatment with conventional and non-conventional anti-parasitic drugs failed to clear parasitosis, we performed targeted metagenomics to observe the bacterial composition of the patient’s gut microbiota (GM), and to evaluate fecal microbiota transplantation (FMT) as a potential treatment option. We found that C. parvum infection led to significant shifts in GM bacterial composition in our patient, with consequent shifts in predicted intestinal functional signatures consistent with a state of persistent inflammation. This, combined with the patient’s poor prognosis and increasing parasitic burden despite many rounds of anti-parasitic drug treatments, made the patient a potential candidate for an experimental FMT procedure. Unfortunately, given the many comorbidities that were precipitated by the patient’s immunodeficiency and chronic C. parvum infection, FMT was postponed in favor of more urgently necessary liver and bone marrow transplants. Tragically, after the first liver transplant failed, the patient lost his life before undergoing FMT and a second liver transplant. With this case report, we present the first description of how cryptosporidiosis can shape the gut microbiota of a pediatric patient with severe immunodeficiency. Finally, we discuss how both our results and the current scientific literature suggest that GM modulations, either by probiotics or FMT, can become novel treatment options for chronic Cryptosporidium infection and its consequent complications, especially in those patients who do not respond to the currently available anti-parasitic therapies.

Current medication therapy for leishmaniasis and trypanosomiasis remains a major challenge due to its limited efficacy, significant adverse effects, and inaccessibility. Consequently, locating affordable and effective medications is a pressing concern. Because of their easy-to-understand structure and high functionalization potential, chalcones are promising candidates for use as bioactive agents. Thirteen synthetic ligustrazine-containing chalcones were evaluated for their ability to inhibit the growth of leishmaniasis and trypanosomiasis in etiologic agents. The tetramethylpyrazine (TMP) analogue ligustrazine was chosen as the central moiety for the synthesis of these chalcone compounds. The most effective compound (EC50 = 2.59 µM) was the chalcone derivative 2c, which featured a pyrazin-2-yl amino on the ketone ring and a methyl substitution. Multiple actions were observed for certain derivatives, including 1c, 2a–c, 4b, and 5b, against all strains tested. Eflornithine served as a positive control, and three ligustrazine-based chalcone derivatives, including 1c, 2c, and 4b, had a higher relative potency. Compounds 1c and 2c are particularly efficacious; even more potent than the positive control, they are therefore promising candidates for the treatment of trypanosomiasis and leishmaniasis. [ABSTRACT FROM AUTHOR]

This article investigates the occurrence and distribution of parasitic protozoa of Iranian freshwater fishes (both farmed and wild). Our search shows 26 known parasitic protozoan species were recorded from 52 freshwater fish species across different ecoregions of Iran. Most of these fish are edible. While none of the identified protozoan parasites are of zoonotic importance, our study does not exclude presence of zoonotic species in Iranian fishes. Present data suggest the northern and western regions of the country are the main macrohabitat of protozoa (35 parasitic records reported), with the greatest concentration of parasitic protozoa occurring in the Urmia basin in Iran's northwest. The clustered distribution pattern of protozoa among freshwater fish was also more evident in the northern and western parts of the country. The gills and skin were the most infected microhabitats for parasitic protozoa. The highest number of parasites was observed in the fish family Cyprinidae with nine species found in the native fish, Capoeta capoeta. The most diverse host range was observed in the holotrich ciliate, Ichthyophthirius multifiliis isolated from 46 cyprinid species in 39 different locations. However, due to the great richness of fish and extreme habitat diversity, parts of the parasite fauna of Iranian freshwater fish are still poorly understood. Furthermore, current and future changes in climate and environmental parameters, and anthropogenic interventions are likely to affect fish hosts and their parasites. [ABSTRACT FROM AUTHOR]

The association between the parasitic illnesses and the consumption of contaminated water has been largely reported. However, there is still a lack of studies investigating the extent of parasitic contamination in water in Morocco. This is the first study in Morocco that aimed at assessing the presence of protozoan parasites, namely Cryptosporidium spp., Giardia duodenalis, and Toxoplasma gondii, in drinking water consumed in the region of Marrakech.Samples processing was performed by membrane filtration and qPCR detection. A total of 104 drinking water samples (tap water, well, and spring waters) was collected between 2016 and 2020.The analysis revealed an overall protozoa contamination rate of 67.3% (70/104), of which 35 samples were positive for Giardia duodenalis, 18 for Toxoplasma gondii, and 17 for both parasites, whereas no sample was positive for Cryptosporidium spp.This first study showed that drinking water in the region of Marrakech contained parasites which could represent a risk for consumers. For a better understanding and estimation of the risk encountered by local inhabitants, further studies concerned with (oo)cyst viability, infectivity, and genotype identification need to be performed.

Introduction: Parasitic intestinal diseases caused by protozoans and helminths are important causes of gastrointestinal disturbances which are responsible for a high burden of morbidity and mortality as they are detrimental not only to paediatric and immunocompromised patients but also to adults and immunocompetent patients. Hence, it is important to know their burden in a geographical area to develop adequate control measures. Aim: To estimate the prevalence of various intestinal parasitic infections in symptomatic patients based on age, gender and immune status in a tertiary care hospital, New Delhi, India. Materials and Methods: A retrospective analysis of 1431 stool samples which were received in the Department of Microbiology VMMC and Safdarjung Hospital, New Delhi, from April 2019 to March 2020 was done. All samples were subjected to macroscopic and microscopic examinations and underwent, wet mount (saline and iodine) examination and Modified Ziehl-Neelson staining. Data was analysed using Statistical Package for Social Sciences (SPSS) software version 21.0 with two-tailed Chi-square test wherever applicable. Results: Males (61.36%) outnumbered females. Prevalence of parasites was more in adults (7.76%) than in children. Higher positivity (7.6%) was observed in immunocompetent individuals as compared to immunocompromised. Overall predominance of protozoans (98.26%) over helminths was noted. Entamoeba histolytica/dispar group (43.60%) was found to be the most prevalent parasite. Entamoeba histolytica/dispar group (5.64%) and Cryptosporidium spp. (5.94%) were most common amongst immunocompetent and immunocompromised patients respectively. Conclusion: A higher prevalence of intestinal parasitic infections in immunocompetent symptomatic patients was observed, requiring attention of clinicians as they are no longer limited majorly to immunocompromised patients. As no effective vaccine is available for these infections, so appropriate laboratory methods, microbiological expertise, proper sanitation measures, availability of potable water supply and properly cooked food are essential to control parasitic intestinal diseases.

Protozoan parasites, such as Plasmodium, Leishmania, Toxoplasma, Cryptosporidium, and Trypanosoma, are causative agents of health-threatening diseases in both humans and animals, leading to significant health risks and socioeconomic losses globally. The development of effective therapeutic and prevention strategies for protozoan-caused diseases requires a full understanding of the pathogenesis and protective events occurring in infected hosts. Interferons (IFNs) are a family of cytokines with diverse biological effects in host antimicrobial defense and disease pathogenesis, including protozoan parasite infection. Type II IFN (IFN-γ) has been widely recognized as the essential defense cytokine in intracellular protozoan parasite infection, whereas recent studies also revealed the production and distinct function of type I and III IFNs in host defense against these parasites. Decoding the complex network of the IFN family in host–parasite interaction is critical for exploring potential new therapeutic strategies against intracellular protozoan parasite infection. Here, we review the complex effects of IFNs on the host defense against intracellular protozoan parasites and the crosstalk between distinct types of IFN signaling during infections. [ABSTRACT FROM AUTHOR]

The complement system exerts crucial functions both in innate immune responses and adaptive humoral immunity. This pivotal system plays a major role dealing with pathogen invasions including protozoan parasites. Different pathogens including parasites have developed sophisticated strategies to defend themselves against complement killing. Some of these strategies include the employment, mimicking or inhibition of host's complement regulatory proteins, leading to complement evasion. Therefore, parasites are proven to use the manipulation of the complement system to assist them during infection and persistence. Herein, we attempt to study the interaction´s mechanisms of some prominent infectious protozoan parasites including Plasmodium, Toxoplasma, Trypanosoma, and Leishmania dealing with the complement system. Moreover, several crucial proteins that are expressed, recruited or hijacked by parasites and are involved in the modulation of the host´s complement system are selected and their role for efficient complement killing or lysis evasion is discussed. In addition, parasite's complement regulatory proteins appear as plausible therapeutic and vaccine targets in protozoan parasitic infections. Accordingly, we also suggest some perspectives and insights useful in guiding future investigations. [ABSTRACT FROM AUTHOR]

Protozoan parasites are responsible for several harmful and widespread human diseases that cause high morbidity and mortality. Currently available treatments have serious limitations due to poor efficiency, strong adverse effects, and high cost. Hence, the identification of new targets and the development of specific drug therapies against parasitic diseases are urgent needs. Heat shock protein 90 (HSP90) is an ATP-dependent molecular chaperone that plays a key role in parasite survival during the various differentiation stages, spread over the vector insect and the human host, which they undergo during their life cycle. The N-terminal domain (NTD) of HSP90, containing the main determinants for ATPase activity, represents the most druggable domain for inhibitor targeting. The molecules investigated on parasite HSP90 are mainly developed from known inhibitors of the human counterpart, and they have strong limitations due to selectivity issues, accounting for the high conservation of the ATP-binding site between the parasite and human proteins. The current review highlights the recent structural progress made to support the rational design of new molecules able to effectively block the chaperone activity of parasite HSP90. [ABSTRACT FROM AUTHOR]

The protozoan parasite Leishmania, responsible for leishmaniasis, is one of the few aerobic organisms that cannot synthesize the essential molecule heme. Therefore, it has developed specialized pathways to scavenge it from its host. In recent years, some proteins involved in the import of heme, such as LHR1 and LFLVCRB, have been identified, but relevant aspects regarding the process remain unknown. Here, we characterized the kinetics of the uptake of the heme analogue Zn(II) Mesoporphyrin IX (ZnMP) in Leishmania major promastigotes as a model of a parasite causing cutaneous leishmaniasis with special focus on the force that drives the process. We found that ZnMP uptake is an active, inducible, and pH-dependent process that does not require a plasma membrane proton gradient but requires the presence of the monovalent cations Na+ and/or K+. In addition, we demonstrated that this parasite can efflux this porphyrin against a concentration gradient. We also found that ZnMP uptake differs among different dermotropic or viscerotropic Leishmania species and does not correlate with LHR1 or LFLVCRB expression levels. Finally, we showed that these transporters have only partially overlapping functions. Altogether, these findings contribute to a deeper understanding of an important process in the biology of this parasite. [ABSTRACT FROM AUTHOR]