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Giardia lamblia is an important protozoan cause of diarrheal disease worldwide, delayed development and cognitive impairment in children in low- and middle-income countries, and protracted post-infectious syndromes in developed regions. G. lamblia resides in the lumen and at the epithelial surface of the proximal small intestine but is not mucosa invasive. The protozoan parasite is genetically diverse with significant genome differences across strains and assemblages. Animal models, particularly murine models, have been instrumental in defining mechanisms of host defense against G. lamblia, but mice cannot be readily infected with most human pathogenic strains. Antibiotic pretreatment can increase susceptibility, suggesting that the normal microbiota plays a role in controlling G. lamblia infection in mice, but the broader implications on susceptibility to diverse strains are not known. Here, we have used gnotobiotic mice to demonstrate that robust intestinal infection can be achieved for a broad set of human-pathogenic strains of the genetic assemblages A and B. Furthermore, gnotobiotic mice were able to eradicate infection with a similar kinetics to conventional mice after trophozoite challenge. Germ-free mice could also be effectively immunized by the mucosal route with a protective antigen, α1-giardin, in a manner dependent on CD4 T cells. These results indicate that the gnotobiotic mouse model is powerful for investigating acquired host defenses in giardiasis, as the mice are broadly susceptible to diverse G. lamblia strains yet display no apparent defects in mucosal immunity needed for controlling and eradicating this lumen-dwelling pathogen.

Introduction: Post-translational modifications of proteins provide cellular physiology with a broad range of adaptability to the external environment flexibly and rapidly. In the case of the protozoan parasite Giardia lamblia , the study of these modifications has gained relevance in recent years, mainly focusing on methylation and deacetylation of proteins. This study investigates the significance of acetylation in this protozoan parasite. Methods: This study explores the role of acetylation in G. lamblia through a combination of immunofluorescence assays, manipulation of acetyltransferase enzymes, and the use of garcinol, an acetylation inhibitor, during the growth phase. Results: The acetylation of histone marks H3K9 and H3K27 occurs during growth and is followed by deacetylation during encystation. Transfections modifying acetyltransferase activity induced a latent cellular state, underscoring the importance of protein acetylation for parasite survival. Garcinol treatment during growth caused significant morphological changes, including plasma membrane blebbing and apoptotic-like bodies. Immunofluorescence revealed these bodies contained α-tubulin/acetylated α-tubulin and reactive oxygen species. Flow cytometry and Annexin V staining indicated early apoptosis within 24 hours of treatment. Additionally, garcinol led to the deacetylation of H3K9 and H3K27, with redistribution of tubulin and acetylated tubulin from microtubules to the cytosol. Significantly, garcinol prevented parasite recrudescence after treatment withdrawal. Discussion: These results demonstrate that acetylation is essential for trophozoite survival and highlight the natural histone acetyltransferase inhibitor garcinol as a potential candidate for drug development against giardiasis, considering its giardicidal activity. [ABSTRACT FROM AUTHOR]

Background: Intestinal parasitic infections (IPIs) are a major health problem among the Karen hill tribe in Thailand. This study aimed to evaluate the effect of an integrated intervention program of drug therapy combined with water, sanitation, and hygiene (WASH) education on reinfection with IPIs among the Karen hill tribe in an endemic area of northern Thailand. Methods: A quasi-experimental study was conducted in two Karen villages, involving 691 residents, in Omkoi District, Chiang Mai Province; one village was designated as the intervention group and the other as the control group. Baseline information was collected regarding the infections and participants' knowledge, attitudes, and practices (KAP) related to prevention and control of IPIs. Detection of benzimidazole resistance linked to the beta-tubulin gene mutation in soil-transmitted helminths (STH) was performed using polymerase chain reaction (PCR) amplification and DNA sequencing. Mass drug administration (MDA) with albendazole was applied to both groups. The intervention group received WASH education, whereas the control group did not. Follow-up assessments were conducted at 3 and 6 months. Results: Baseline data revealed a 36.0% (123/342) prevalence of IPIs in the intervention group and 36.8% (96/261) in the control group. The most common helminth and pathogenic protozoan were Trichuris trichiura and Giardia lamblia, respectively. No non-synonymous mutations in the beta-tubulin gene were found. Post-intervention at 3-month and 6-month follow-ups revealed that the prevalence of IPIs in the intervention group was significantly decreased to 23.6% (P = 0.002) and 23.1% (P = 0.002), and the prevalence of pathogenic IPIs was reduced from 9.4% to 3.9% (P = 0.013) and 2.4% (P = 0.002), respectively. In contrast, no significant changes in the prevalence of IPIs were observed in the control group. The intervention group showed significant improvements in KAP scores, which were significantly higher than those in the control group. Conclusions: MDA alone is not effective for controlling IPIs among the Karen people due to rapid reinfection related to behavioral factors and socioeconomic conditions. We demonstrated for the first time that integration of WASH education increased KAP scores and consequently significantly reduced IPI reinfection among the Karen hill tribe in northern Thailand. [ABSTRACT FROM AUTHOR]

In this study, n-butyl and iso-butyl quinoxaline-7-carboxylate-1,4-di-N-oxide derivatives were evaluated in vitro against Giardia lamblia (G. lamblia), Trichomonas vaginalis (T. vaginalis), and Entamoeba histolytica (E. histolytica). The potential mechanism of action determination was approached by in silico analysis on G. lamblia and T. vaginalis triosephosphate isomerase (GlTIM and TvTIM, respectively), and on E. histolytica thioredoxin reductase (EhTrxR). Enzyme inactivation assays were performed on recombinant GlTIM and EhTrxR. Compound T-167 showed the best giardicidal activity (IC50 = 25.53 nM) and the highest inactivation efficiency against GlTIM without significantly perturbing its human homolog. Compounds T-142 and T-143 showed the best amoebicidal (IC50 = 9.20 nM) and trichomonacidal (IC50 = 45.20 nM) activity, respectively. Additionally, T-143 had a high activity as giardicial (IC50 = 29.13 nM) and amoebicidal (IC50 = 15.14 nM), proposing it as a broad-spectrum antiparasitic agent. Compounds T-145, and T-161 were the best EhTrxR inhibitors with IC50 of 16 µM, and 18 µM, respectively. [ABSTRACT FROM AUTHOR]

Diseases caused by intestinal parasites such as protozoa represent a worldwide problem mainly for developing countries. From morbidity in different groups of people to cases of mortality in children and/or immunocompromised patients. In addition to the above, there is growing resistance to the drugs used in the treatment of these diseases, as well as undesirable side effects in patients. Therefore, there is an interest in the search for new alternatives for the base and/or development of new drugs with antiparasitic activities without harmful effects. In this sense, natural products offer to be a diverse source of compounds with biological activities. In this work, we describe the isolation and elucidation by 1D and 2D NMR spectroscopy of three cyclopeptides obtained from seeds of A. diversifolia Saff.: cherimolacyclopeptide D (1), squamin D (2), and squamin C (3). The fractions enriched in cyclopeptides, as well as a pure compound (1), showed antiprotozoal activity against E. hystolitica Schaudinn and Giardia lamblia Kunstler in vitro assays, with values of IC50 = 3.49 and 5.39 μg mL−1, respectively. The molecular docking study revealed that 1 has a strong interaction with targets used, including aldose reductase and PFOR enzymes. The antiprotozoal activity of cherimolacyclopeptide D is reported for the first time in this study. [ABSTRACT FROM AUTHOR]

Anemia is a health problem of concern among schoolchildren in underprivileged rural regions, where recurrent parasitic infections are common. A cross-sectional study was conducted in 229 schoolchildren in rural highland Bolivia in the department of La Paz, an area with a high prevalence of protozoan and helminth infections, to determine the types and mechanisms of anemia. A substantial proportion of children (40.2%) were found to be anemic based on hemoglobin measurements. No associations were found between low hemoglobin levels and helminth or protozoan infections when evaluating infectious causes of anemia, nor with Giardia lamblia or Blastocystis hominis, which are associated with iron deficiency and nutrient malabsorption and were highly prevalent in this study. The significant association between anemia and hypochromia suggests iron deficiency, aligned with low hemoglobin levels. A total of 39 out of 150 children (26%) had markers consistent with iron deficiency anemia (IDA), 26 out of 127 children (20%) met the criteria for anemia of inflammation (AI). Furthermore, 12 of the 127 tested children (9.4%) met the criteria for mixed AI with IDA according to the soluble transferrin receptor (sTfR)/log ferritin levels, which increased significantly due to overall infections by Hymenolepis nana and Ascaris lumbricoides helminths. The findings highlight the need for integrated public health interventions to address iron nutrition and parasitic infections to effectively prevent anemia in this vulnerable population. [ABSTRACT FROM AUTHOR]

Purpose: Intestinal protozoan parasites among Asian schoolchildren are a subject of concern due to their prevalence and potential health impact. Understanding and addressing this issue is crucial for public health in the region. Methods: We conducted a comprehensive search for articles published up to December 2023 across four databases, including Scopus, PubMed, ProQuest, and Web of Science. To estimate the combined prevalence, a random-effects model with a 95% confidence interval (CI) was applied, and the statistical analysis was performed using meta-analysis packages in R version (3.6.1). This study is registered with PROSPERO (CRD42023481146). Results: Among 131 eligible articles, the prevalence of intestinal protozoan parasites was 0.208 (95% CI = 0.180–0.238). Lebanon and Tajikistan had the highest country-level prevalence at 0.851 and 0.836, respectively, with Giardia duodenalis being the most prevalent species at 0.082. Conclusion: In summary, our study highlights the urgent public health issue of protozoan parasites among Asian schoolchildren due to poor sanitation and water quality. Immediate interventions are essential, considering climate and socioeconomic factors, to combat these infections and improve overall health. [ABSTRACT FROM AUTHOR]

Background: Giardia lamblia is an intestinal protozoan estimated to cause ~200 million symptomatic infections annually, mainly in children in low- and middle-income countries associated with intestinal damage, increased permeability, and malabsorption. Methods and results: We describe here the epidemiology, incidence, clinical characteristics, and risk factors of acute gastroenteritis episodes (AGE) with G. lamblia detection (GAGE) using a birth cohort of 443 Nicaraguan children followed weekly until 36 months of life. From June 2017 to July 2021, 1385 AGE samples were tested by qPCR. G. lamblia was detected in 104 (7.5%) of AGE episodes. In all, 69 (15.6%) children experienced at least one GAGE episode, and 25 of them (36.2%) experienced more than one episode. The incidence rate of the first episode of GAGE was 6.8/100 child-years (95% CI, 4.5–9.1). During GAGE, bloody stools, vomiting, and fever were uncommon, and children were less likely to be treated at a primary care clinic, suggesting that GAGE is typically mild and most cases did not receive medical attention, which could facilitate higher parasite loads with increased possibilities of establishing chronic carriage. GAGE was more common in children 12–24 months of age (13.9/100 child-years [95% CI, 10.7–17.1]) as compared to other age groups. In our birth-cohort, children living in a home with an indoor toilet (aHR, 0.52 [95%CI, 0.29–0.92]), and being breastfed in the first year of life (aHR: 0.10 [95%IC, 0.02, 0.57]) had a lower incidence of GAGE. In contrast, being breastfed for ≤ 6 months was associated with a higher incidence if the children were living in houses without indoor toilets and earthen floors (HR, 7.79 [95% CI, 2.07, 29.3]). Conclusion: Taken together, GAGE is more frequent under poor household conditions. However, breastfeeding significantly reduces the incidence of GAGE in those children. Author summary: G. lamblia is an intestinal protozoan having mixed associations with acute gastroenteritis, yet positive associations with impaired child growth. G. lamblia infects millions of people worldwide annually and is more prevalent in children residing in low- and middle-income countries, making it an important target for prevention and control efforts. Although G. lamblia carriage is widespread, its epidemiology, natural history, genetic diversity, host genetic susceptibility factors, and post-infectious sequelae in children are still under investigation. As no human vaccine is available, understanding its risk factors in an endemic country is the first step for reducing the burden and mitigating its sequelae. In a birth cohort in León, Nicaragua, we found that children living in households with poor sanitation conditions are at higher risk of AGE with G. lamblia detection (GAGE), but prolonged duration of breastfeeding despite these conditions may reduce the risk in these children. The study highlights the importance of breastfeeding and the need to better understand how G. lamblia contributes to the global burden of disease in children, and the mechanisms of host protection to guide future interventions. [ABSTRACT FROM AUTHOR]

Background: Giardia duodenalis is a common enteric protozoan parasite that is categorized into eight assemblages (A–H). In particular, assemblages A and B are zoonotic, capable of infecting both humans and animals worldwide, resulting in significant economic losses and public health challenges in epidemic regions. Thus, the development of rapid, accurate and non-laboratory-based diagnostic methods for infected animals is crucial for the effective prevention and control of giardiasis. Recent advancements in clustered, regularly interspaced, short palindromic repeats (CRISPR) and CRISPR-associated (Cas) protein (Cas12a) systems allow promising avenues for nucleic acid detection, characterized by their high flexibility, sensitivity and specificity. Methods: Combined recombinase polymerase amplification and CRISPR/Cas12a systems were combined and used as end-point diagnostic methods (termed REPORT) to detect G. duodenalis assemblage A and B. The diagnostic results can be observed by fluorescence readouts with the naked eye under blue light or colorimetric signals using a lateral flow strip (LFS). Results: The limit of detection (LOD) of the REPORT‑based G. duodenalis assemblage A detection was 2.04 CFU/ml and 10 trophozoites per gram (TPG), and the LOD of assemblage B was 1.1 CFU/ml and 10 cysts per gram (CPG). The REPORT‑based G. duodenalis assemblage A and assemblage B detection methods have strong specificity and no cross-reactivity with other assemblages of G. duodenalis or common enteric parasitic protozoa and have excellent performance in clinical sample detection. Conclusions: This study presents a novel strategy for the direct identification of G. duodenalis assemblages A and B, requiring neither highly trained personnel nor costly specialized equipment. [ABSTRACT FROM AUTHOR]

Introduction: Giardia lamblia is a flagellated protozoan parasite causing giardiasis, a common intestinal infection characterized by diarrhea, abdominal cramps, and nausea. Treatments employed to combat this parasitic infection have remained unchanged for the past 40 years, leading to the emergence of resistant strains and prompting the search for new therapeutic agents. Methods: This study investigated the cytotoxic effects of ivermectin (IVM) on G. lamblia trophozoites. We conducted dose-response experiments to assess IVM-induced cytotoxicity. We utilized various biochemical and ultrastructural analyses to explore the underlying mechanisms of cell death, including reactive oxygen species (ROS) production, DNA fragmentation, cell cycle arrest, and apoptosis markers. Results: Our findings demonstrate that IVM induces dose-dependent cytotoxicity and triggers cell death pathways. We found that IVM treatment generates elevated levels of reactive oxygen species (ROS), DNA fragmentation, and arrests of trophozoites in the cell cycle's S phase. Additionally, ultrastructural analysis reveals morphological alterations consistent with apoptosis, such as cytoplasmic vacuolization, chromatin condensation, and tubulin distribution. Discussion: The insights gained from this study may contribute to developing new therapeutic strategies against giardiasis, addressing the challenge posed by drug-resistant strains. [ABSTRACT FROM AUTHOR]

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.

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.

Myeloid leukemia factor 1 (Mlf1) was identified as a proto-oncoprotein that affects hematopoietic differentiation in humans. However, its cellular function remains elusive, spanning roles from cell cycle regulation to modulation of protein aggregate formation and participation in ciliogenesis. Given that structurally conserved homologs of Mlf1 can be found across the eukaryotic tree of life, we decided to characterize its cellular role underlying this phenotypic pleiotropy. Using a model of the unicellular eukaryote Giardia intestinalis, we demonstrate that its Mlf1 homolog (GiMlf) mainly localizes to two types of cytosolic foci: microtubular structures, where it interacts with Hsp40, and ubiquitin-rich, membraneless compartments, found adjacent to mitochondrion-related organelles known as mitosomes, containing the 26S proteasome regulatory subunit 4. Upon cellular stress, GiMlf either relocates to the affected compartment or disperses across the cytoplasm, subsequently accumulating into enlarged foci during the recovery phase. In vitro assays suggest that GiMlf can be recruited to membranes through its affinity for signaling phospholipids. Importantly, cytosolic foci diminish in the gimlf knockout strain, which exhibits extensive proteomic changes indicative of compromised proteostasis. Consistent with data from other cellular systems, we propose that Mlf acts in the response to proteotoxic stress by mediating the formation of function-specific foci for protein folding and degradation. Author summary: Giardia intestinalis, a widespread intestinal parasite, has emerged as a valuable model for studying eukaryotic cell biology and host-pathogen interactions. Our study focuses on the Myeloid Leukemia Factor (Mlf) homolog in Giardia (GiMlf), an evolutionarily conserved protein with diverse cellular functions. We demonstrate that GiMlf plays a crucial role in proteostasis by forming two types of cytosolic foci: one associated with microtubular structures and Hsp40, and another comprising ubiquitin-rich, membraneless compartments near mitosomes, containing the 26S proteasome regulatory subunit. Upon cellular stress, GiMlf dynamically relocates, suggesting its involvement in the stress response. Notably, GiMlf knockout leads to extensive proteomic changes and altered encystation rates, indicating its importance in Giardia's life cycle and stress adaptation. Our findings provide insights into how Giardia, and potentially other eukaryotes, maintain proteostasis under various environmental conditions. This research enhances our understanding of fundamental parasite biology and stress response mechanisms, which are critical for pathogen survival in diverse host environments. [ABSTRACT FROM AUTHOR]

Lymphangioleiomyomatosis (LAM) is a rare genetic lung disease. Unfortunately, treatment with the mTORC1 inhibitor Rapamycin only slows disease progression, and incomplete responses are common. Thus, there remains an urgent need to identify new targets for the development of curative LAM treatments. Nitazoxanide (NTZ) is an orally bioavailable antiprotozoal small molecule drug approved for the treatment of diarrhea caused by Giardia lamblia or Cryptosporidium parvum in children and adults, with a demonstrated mTORC1 inhibitory effect in several human cell lines. NTZ's excellent safety profile characterized by its more than 20 years of clinical use makes it a promising candidate for repurposing. Our rationale for this study was to further investigate NTZ's effect using in vitro and in vivo LAM models and to elucidate the underlying molecular mechanism beyond mTORC1 inhibition. For this purpose, we investigated cell proliferation, cell viability, and changes in protein phosphorylation and expression in primary human cell cultures derived from LAM lung samples before translating our results into a syngeneic mouse model utilizing Tsc2-null cells. NTZ reduced cell growth for all tested cell lines at a dose of about 30 µM. Lower doses than that had no effect on cell viability, but doses above 45 µM lowered the viability by about 10 to 15% compared to control. Interestingly, our western blot revealed no inhibition of mTORC1 and only a mild effect on active ß-Catenin. Instead, NTZ had a pronounced effect on reducing pAkt. In the mouse model, prophylactic NTZ treatment via the intraperitoneal and oral routes had some effects on reducing lung lesions and improving body weight retention, but the results remain inconclusive. [ABSTRACT FROM AUTHOR]

Background : Diplomonads are anaerobic flagellates classified within Metamonada. They contain both host-associated commensals and parasites that reside in the intestinal tracts of animals, including humans (e.g., Giardia intestinalis), as well as free-living representatives that inhabit freshwater and marine anoxic sediments (e.g., Hexamita inflata). The evolutionary trajectories within this group are particularly unusual as the free-living taxa appear to be nested within a clade of host-associated species, suggesting a reversal from host-dependence to a secondarily free-living lifestyle. This is thought to be an exceedingly rare event as parasites often lose genes for metabolic pathways that are essential to a free-living life strategy, as they become increasingly reliant on their host for nutrients and metabolites. To revert to a free-living lifestyle would require the reconstruction of numerous metabolic pathways. All previous studies of diplomonad evolution suffered from either low taxon sampling, low gene sampling, or both, especially among free-living diplomonads, which has weakened the phylogenetic resolution and hindered evolutionary insights into this fascinating transition. Results: We sequenced transcriptomes from 1 host-associated and 13 free-living diplomonad isolates; expanding the genome scale data sampling for diplomonads by roughly threefold. Phylogenomic analyses clearly show that free-living diplomonads form several branches nested within endobiotic species. Moreover, the phylogenetic distribution of genes related to an endobiotic lifestyle suggest their acquisition at the root of diplomonads, while traces of these genes have been identified in free-living diplomonads as well. Based on these results, we propose an evolutionary scenario of ancestral and derived lifestyle transitions across diplomonads. Conclusions: Free-living taxa form several clades nested within endobiotic taxa in our phylogenomic analyses, implying multiple transitions between free-living and endobiotic lifestyles. The evolutionary history of numerous virulence factors corroborates the inference of an endobiotic ancestry of diplomonads, suggesting that there have been several reversals to a free-living lifestyle. Regaining host independence may have been facilitated by a subset of laterally transferred genes. We conclude that the extant diversity of diplomonads has evolved from a non-specialized endobiont, with some taxa becoming highly specialized parasites, others becoming free-living, and some becoming capable of both free-living and endobiotic lifestyles. Highlights: • Expanding the taxon sampling of phylogenomic analyses for diplomonads by sequencing transcriptomes of 13 free-living isolates and one endobiotic isolate. • Constructing a well-resolved evolutionary framework for exploring lifestyle adaptation in diplomonads using a phylogenomic approach. • Identifying genetic signatures of parasitic ancestry (variant-specific surface proteins, virulence factors) in free-living diplomonads. [ABSTRACT FROM AUTHOR]

Background: The urease-producing Helicobacter pylori increase the likelihood that pathogenic intestinal protozoa will use the stomach's increased hydrogen potential to propagate the disease. Coinfections exacerbate the onset and severity of gastrointestinal symptoms. This study aimed to assess the prevalence of intestinal parasites/Helicobacter pylori coinfection and contributing factors in patients with gastrointestinal symptoms at Addis Zemen Primary Hospital, Northwest Ethiopia. Methods: From April to July 2023, patients with gastrointestinal problems participated in a cross-sectional study carried out in a hospital. To collect the clinical and sociodemographic data, a questionnaire was employed. Intestinal parasites and Helicobacter pylori were detected using the saline stool wet mount and Helicobacter pylori stool antigen tests, respectively. SPSS version 20 was used to analyze the data and variables with p-values < 0.05 were considered statistically significant. Result: The study included 384 participants in total, of which 47.3% (182/384) were farmers and 50.3% (193/384) were women. Of the study subjects, 69/384 or approximately 18%, had intestinal parasitic infections. In 12% of cases (46/384), Helicobacter pylori were detected. A coinfection of Helicobacter pylori and intestinal parasites was found in 5.5% (21/384) of the subjects. Multiple logistic regression revealed increased risk of coinfection of Helicobacter pylori and intestinal parasites in patients who drink surface water (AOR: 10.7, p = 0.03) family history of Helicobacter pylori (AOR: 3.3, p = 0.024) and those with untrimmed fingers (AOR: 4.9, p = 0.031). Conclusions: Giardia lamblia and Entamoeba histolytica/dispar/ moshkovskii/ bangladeshi complex are the most common protozoans that cause coinfection with Helicobacter pylori. Drinking surface water, family history of Helicobacter pylori and untrimmed fingers are the contributing factors to intestinal parasites/Helicobacter pylori coinfection. [ABSTRACT FROM AUTHOR]

Giardia doudenalis (lamblia, intestinalis) is a protozoan parasite that inhabits the lumen of the upper small intestine of vertebrates, causing chronic abdominal pains and severe diarrhea, symptoms of giardiasis, a persistent and recurrent infection. This characteristic is mainly due to the presence of membrane variant-specific surface proteins (VSPs) that give this parasite the ability to successively infect the host through antigenic variation. Using high-resolution scanning microscopy (HR-SM), we observed the presence, formation, and extension of tunneling-nanotube-like surface structures in Giardia, especially following parasite challenges with VSP antibodies. They were seen all over the parasite surface, both in vitro and in vivo, showing that G. duodenalis nanotube formation occurs in complex environments such as the gut. In addition, we also observed that some of these nanotubes displayed a periodic strangulation that produces 100 nm vesicles that seemed to be released in a process similar to that previously observed in Trypanosoma brucei. The presence of nanotube-like structures in G. duodenalis highlights yet another strategy of cellular communication utilized by these parasites, whether between themselves or with the host cell. [ABSTRACT FROM AUTHOR]

Bacillus spp. are foodborne pathogens important in food contamination, especially in dried milk products. They are associated with foodborne outbreaks, spoilage of dairy products (sweet curdling and bitterness), and bovine mastitis. This study investigated the contamination rate of Bacillus spp. in a total of 105 samples of the dried milk products kishk, Cerelac and whole milk powder, marketed in Assiut city, Upper Egypt. The identification of the Bacillus spp. strains was completed using conventional biochemical methods and PCR protocols for the confirmation of isolates. Bacillus spp. were isolated from 57 out of 105 (54%) [95% CI: 44.6-64.0%] of the examined samples at levels of up to 7.7 log10 cfu/g. The prevalence of Bacillus spp. was significantly (P<0.05) higher in kishk (74%) than in the other food categories with a mean count of 4.04±0.49 log10 cfu/g. Regarding the species of Bacillus detected, 39.05% of the examined samples contained B. cereus, 8.57% contained B. subtilis, 2.86%: B. pumilus, another 2.86%: B. megatrium and 0.95%: B. licheniformis. B. cereus was isolated from 18 (51.43%) of the kishk samples, 9 (25.71%) of the Cerelac samples and 14 (40%) of the whole milk powder samples. Out of the 41 B. cereus strains previously identified by biochemical tests, 19 (46.34%) isolates were also confirmed using PCR (61.11%, 42.86% and 22.22% for kishk, whole milk powder and Cerelac, respectively). In conclusion, incorporating preventive measures to reduce bacterial contamination in the Egyptian dairy environment are warranted in order to avoid the contamination of milk products with these life-threatening pathogens. [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]

Giardia lamblia, the cause of giardiasis, significantly impacts patients with metabolic disorders related to insulin resistance (IR). Both giardiasis and metabolic disorders share elements such as chronic inflammation and intestinal dysbiosis, which substantially affect the metabolic and cytokine profiles of patients. This review discusses the mechanisms of virulence of G. lamblia, its influence on the immune system, and its association with metabolic disorders. The review aims to show how G. lamblia invasion acts on the immune system and the glucose and lipid metabolism. Key findings reveal that G. lamblia infection, by disrupting intestinal permeability, alters microbiota composition and immune responses, potentially impairing metabolic status. Future research should focus on elucidating the specific mechanisms by which G. lamblia influences the metabolism, exploring the long-term consequences of chronic infection, and developing targeted therapeutic strategies that include both parasitic and metabolic aspects. These insights underscore the need for a multidisciplinary approach to the treatment of giardiasis in patients with metabolic disorders. [ABSTRACT FROM AUTHOR]

Background: The Gran Chaco ecoregion is a well-known hotspot of several neglected tropical diseases (NTDs) including Chagas disease, soil-transmitted helminthiasis and multiparasitic infections. Interspecific interactions between parasite species can modify host susceptibility, pathogenesis and transmissibility through immunomodulation. Our objective was to test the association between human co-infection with intestinal parasites and host parasitaemia, infectiousness to the vector and immunological profiles in Trypanosoma cruzi-seropositive individuals residing in an endemic region of the Argentine Chaco. Methods: We conducted a cross-sectional serological survey for T. cruzi infection along with an intestinal parasite survey in two adjacent rural villages. Each participant was tested for T. cruzi and Strongyloides stercoralis infection by serodiagnosis, and by coprological tests for intestinal parasite detection. Trypanosoma cruzi bloodstream parasite load was determined by quantitative PCR (qPCR), host infectiousness by artificial xenodiagnosis and serum human cytokine levels by flow cytometry. Results: The seroprevalence for T. cruzi was 16.1% and for S. stercoralis 11.5% (n = 87). We found 25.3% of patients with Enterobius vermicularis. The most frequent protozoan parasites were Blastocystis spp. (39.1%), Giardia lamblia (6.9%) and Cryptosporidium spp. (3.4%). Multiparasitism occurred in 36.8% of the examined patients. Co-infection ranged from 6.9% to 8.1% for T. cruzi-seropositive humans simultaneously infected with at least one protozoan or helminth species, respectively. The relative odds of being positive by qPCR or xenodiagnosis (i.e. infectious) of 28 T. cruzi-seropositive patients was eight times higher in people co-infected with at least one helminth species than in patients with no such co-infection. Trypanosoma cruzi parasite load and host infectiousness were positively associated with helminth co-infection in a multiple regression analysis. Interferon-gamma (IFN-γ) response, measured in relation to interleukin (IL)-4 among humans infected with T. cruzi only, was 1.5-fold higher than for T. cruzi-seropositive patients co-infected with helminths. The median concentration of IL-4 was significantly higher in T. cruzi-seropositive patients with a positive qPCR test than in qPCR-negative patients. Conclusions: Our results show a high level of multiparasitism and suggest that co-infection with intestinal helminths increased T. cruzi parasitaemia and upregulated the Th2-type response in the study patients. [ABSTRACT FROM AUTHOR]

Background: Giardiasis, caused by the protozoan parasite Giardia intestinalis, often presents a treatment challenge, particularly in terms of resistance to metronidazole. Despite extensive research, markers for metronidazole resistance have not yet been identified. Methods: This study analysed 28 clinical samples of G. intestinalis from sub-assemblage AII, characterised by varying responses to metronidazole treatment. We focussed on copy number variation (CNV) of the multi-copy flavohemoprotein gene, analysed using digital polymerase chain reaction (dPCR) and next generation sequencing (NGS). Additionally, chromosomal ploidy was tested in 18 of these samples. Flavohemoprotein CNV was also assessed in 17 samples from other sub-assemblages. Results: Analyses revealed variable CNVs of the flavohemoprotein gene among the isolates, with no correlation to clinical metronidazole resistance. Discrepancies in CNVs detected from NGS data were attributed to biases linked to the whole genome amplification. However, dPCR helped to clarify these discrepancies by providing more consistent CNV data. Significant differences in flavohemoprotein CNVs were observed across different G. intestinalis sub-assemblages. Notably, Giardia exhibits a propensity for aneuploidy, contributing to genomic variability within and between sub-assemblages. Conclusions: The complexity of the clinical metronidazole resistance in Giardia is influenced by multiple genetic factors, including CNVs and aneuploidy. No significant differences in the CNV of the flavohemoprotein gene between isolates from metronidazole-resistant and metronidazole-sensitive cases of giardiasis were found, underscoring the need for further research to identify reliable genetic markers for resistance. We demonstrate that dPCR and NGS are robust methods for analysing CNVs and provide cross-validating results, highlighting their utility in the genetic analyses of this parasite. [ABSTRACT FROM AUTHOR]

The current study aimed to detect Giardia lamblia subtypes that infect children under 13 years and determine the association of subtype infection with some risk factors as age, gender, and residence. Triosephosphate isomerase gene from the positive microscopic samples was isolated. To confirm infection and the sensitivity of this gene, DNA in one negative and some positive samples were confirmed when amplified using β-giardin gene primers. From the 66 samples positive by microscopy, 65 tested positive by molecular assay of the triosephosphate isomerase gene with 98.4% sensitivity. While 37/65 (56.9%) represented subtype B, 23/65 (35.4%) represented subtype A, 5 (7.6%) represented A+B, while there was no E subtype. A subtype A in asymptomatic children exceeded 10/25 (40%) compared to 13/40 (32.5%) in symptomatic children. The proportion of B subtype among symptomatic children was higher than that of asymptomatic children (57.5% and 56% respectively). There was no significant difference between the subtype presence and gender, age or residence. Subtype A was higher in the watery diarrhoea samples than in the steatorrhea samples. On the contrary, B subtype was higher in the steatorrhea samples than in watery diarrhoea. Use of the gene triosephosphate isomerase showed a high sensitivity for molecular detection of Giardia spp. Children could be infected with both groups A and B subtypes. The study did not record any presence of subtype E. No significant difference between genders, age groups or place of residence was recorded. Symptoms and diarrhoea were insignificantly associated with subtype B. [ABSTRACT FROM AUTHOR]

Artocapus heterophyllus (jackfruit) is available in abundance in tropical regions. The present study explored the effect of different extraction method i.e. maceration, ultrasonication and both of ultrasonication and maceration on its biological activity (antioxidant, antidiabetes and antibacterial activities) and phytochemical content of Artocarpus heterophyllus (jackfruit) leaves extract. Jackfruit leaves were extracted using ethanol as solvent. The capability of jackfruit leaves extract to scavenge radicals was evaluated using a DPPH (2,2-diphenyl-1-picryl-hydrazyl) assay, β-carotene bleaching assay and reducing power assay. The total phenolic content (TPC) was assessed using the Folin- Ciocalteau method. Its antibacterial activity was assessed using the disc diffusion and microdilution method against Staphylococcus aureus, Bacillus subtilis, Pseudomonas aeruginosa, Salmonella typhimurim and Escherichia coli. The α-glucosidase inhibitor assay was conducted using α-glucosidase from Saccharomyces cerevisiae. The results revealed that ulrasonication method give the highest antioxidant activity in three metode used, i.e. DPPH assay with IC 50 of 42.51 µg/mL, β-carotene bleaching assay of 57.64%, reducing power assay of 919 mg GAE/g. Meanwhile, for α-glucosidase inhibitory assay, ultrasonication method also had the highest value of 83.44% at concentration of 1000 µg/mL. Furthermore, antibacterial assay using disk diffusion method showed ultrasonication had the highest inhibition zone of 7.81 mm at 5 mg/mL against P. aeruginosa , while maceration method had the highest inhibition zone of 8.02 mm at 5 mg/mL against E. coli. These results in accordance with total phenolic content (TPC) of ultrasonication extract which was the highest of 125.06 mg GAE/g. The liquid chromatography-high-resolution mass spectrometry (LC-HRMS) analysis suggested that several bioactive compounds such as phenolic, flavonoid and its derivatives was present in the jackfruit leaves extract. Further analysis using principal component analysis (PCA) revealed that the extraction process affected the metabolite composition of the samples which was confirmed by significant separation of the score plots of extraction methods. The present study revealed that jackfruit leaves is a potent source of natural antibacterial, antioxidants and antidiabetic medicines in functional foods and pharmaceutical industries. [Display omitted] • Different extraction method of jackfruit leaves had been investigated. • Antioxidant, antibacterial and antidiabetic activities of extracts were evaluated. • The ultrasonication method of jackfruit leaves was the most active extracts. • PCA analysis suggested the effect of extraction on the metabolits. [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]

Background: Parasitic infections remain a significant public health concern, particularly in underserved communities where access to healthcare and sanitation is limited. This study aims to investigate the incidence and pathological characteristics of parasitic infections in such communities, providing insights into the prevalence and impact of these infections on public health. Materials and Methods: A cross-sectional study was conducted in underserved communities over a period of one year. A total of 500 participants were randomly selected and screened for parasitic infections through stool and blood samples. The samples were analyzed using microscopy and polymerase chain reaction (PCR) techniques to identify and quantify parasitic species. Pathological examinations were performed to assess the impact of infections on host tissues. Results: The incidence of parasitic infections was found to be 35% among the study population. The most common parasites identified were Ascaris lumbricoides (40%), Giardia lamblia (25%), and Entamoeba histolytica (20%). Pathological analysis revealed that Ascaris lumbricoides caused significant intestinal inflammation in 30% of infected individuals, while Giardia lamblia was associated with malabsorption syndrome in 15% of cases. Entamoeba histolytica was linked to liver abscesses in 10% of the infected participants. Conclusion: Parasitic infections are highly prevalent in underserved communities, with significant pathological impacts on affected individuals.

ABSTRACT: Background: Parasitic infections remain a significant public health concern, particularly in underserved communities where access to healthcare and sanitation is limited. This study aims to investigate the incidence and pathological characteristics of parasitic infections in such communities, providing insights into the prevalence and impact of these infections on public health. Materials and Methods: A cross-sectional study was conducted in underserved communities over a period of one year. A total of 500 participants were randomly selected and screened for parasitic infections through stool and blood samples. The samples were analyzed using microscopy and polymerase chain reaction (PCR) techniques to identify and quantify parasitic species. Pathological examinations were performed to assess the impact of infections on host tissues. Results: The incidence of parasitic infections was found to be 35% among the study population. The most common parasites identified were Ascaris lumbricoides (40%), Giardia lamblia (25%), and Entamoeba histolytica (20%). Pathological analysis revealed that Ascaris lumbricoides caused significant intestinal inflammation in 30% of infected individuals, while Giardia lamblia was associated with malabsorption syndrome in 15% of cases. Entamoeba histolytica was linked to liver abscesses in 10% of the infected participants. Conclusion: Parasitic infections are highly prevalent in underserved communities, with significant pathological impacts on affected individuals. [ABSTRACT FROM AUTHOR]

Understanding cellular ultrastructure is tightly bound to microscopic resolution and the ability to identify individual components at that resolution. Expansion microscopy has revolutionised this topic. Here we present and compare two protocols of ultrastructure expansion microscopy that allow for 4.5-fold mostly isotropic expansion and the use of antibodies, metabolic labelling, and DNA stains to demarcate individual regions such as the endoplasmic reticulum, the nuclei, the peripheral endocytic compartments as well as the ventral disc and the cytoskeleton in Giardia lamblia. We present an optimised, shortened, and modular protocol that can be swiftly adjusted to the investigators needs in this important protozoan model organism.

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.

Giardiasis is among the most prevalent protozoan infections around the globe infecting various species of mammals, birds, reptiles, amphibians, and humans. Among all the species of Giardia, only Giardia lamblia (assemblages A and B) have high zoonotic importance. It is an enteric disease marked by dysentery, abdominal cramps, perfused diarrhea, and anorexia. The giardiasis is majorly controlled by metronidazole, which plays a primary role in the control. The reports of resistance, ecotoxicity, and side effects of this drug necessitate the need for an alternative for the control of this disease; among all the alternatives, phytochemicals are the most promising substance to be used for future anti-giardiasis drug development. Plant preparations containing simple phenolics, alkaloids, saponins, flavonoids, and vitamins have been recently used by researchers. These drugs have been proven effective because of several direct and indirect mechanisms. Simple phenolics easily penetrate the cell of Giardia and disturb energy synthesis, flavonoids destroy the enzymatic process, and alkaloids disturb glycolysis. The vitamins alter the cell energy primarily. Because of these actions, they can be used for control of giardiasis. However, their pharmacological interactions and clinical toxicity studies are needed for their future use. [ABSTRACT FROM AUTHOR]

Background: Giardia intestinalis is a flagellate protozoan that infects both humans and animals. Formation of reactive oxygen species (ROS) during several physiological processes in tissues and cells is indicative of the pathogenesis of different parasitic infections involving Giardia lamblia. Objectives: the current study aimed to assess the potential effects of Punica granatum on interactions of G. intestinalis with the intestinal barrier and its role in ameliorating oxidative stress in a murine model using serum biochemical analysis of both malondialdehyde (MDA) and reduced glutathione (GSH). Methodology: Five experimental groups were involved: Group I included control healthy, group II included infected untreated, group III included infected-treated with metronidazole, group IV included infected-treated with P. granatum, Group V received P. granatum for seven days before and also during induction of infection then regular administration of the drug for an additional seven days. Results: The highest serum levels of MDA (nmol/ml) were expressed in group II 7.03 ± 0.19 followed by group IV that was 5.29 ± 0.62 then group V 4.17 ± 0.28, while group III had 3.58 ± 0.18. On the other side, the highest serum levels of GSH (nmol/ml) were expressed in group V 9.93 ± 0.18, followed by group IV 7.41 ± 0.19 nmol/ml, while in group III was 6.11 ± 0.40. Group II had 5.00 ± 0.15 nmol/ml. There was significant difference between different groups. Conclusions: Decreased expression of MDA in prophylactic and treated groups by P. granatum, moreover increased expression of GSH in group V prophylactic group explains the protective effect of P. granatum against lipid peroxidation and oxidative stress induced by Giardia infection. [ABSTRACT FROM AUTHOR]

Giardiasis is a parasitic disease caused by Giardia lamblia (G. lamblia) that affects people worldwide. Still, few studies report on the immunoregulatory effects of the biomolecules of colostrum during interactions with G. lamblia. This study aimed to assess the concentrations of melatonin and cortisol hormones, the percentage of Treg cells, and the levels of cytokines IL-10 and TGF-β in colostrum from mothers who tested positive for the parasite. This cross-sectional study analyzed colostrum samples from 25 puerperal. The samples were tested using an ELISA to determine if they were seropositive for G. lamblia and the type of antibody present (IgM and IgG). Based on the results, the samples were divided into three groups: a control group (N = 10) with no reaction to either IgM or IgG, a group seropositive for IgG (IgG+/IgM−; N = 8), and a group seropositive for IgM (IgM+/IgG−; N = 7). The concentrations of melatonin and cortisol were measured using the ELISA method. Additionally, cytokines IL-10 and TGF-β and immunophenotyping were analyzed using flow cytometry. In the group that tested positive for IgM anti-G. lamblia, the concentration of melatonin was lower. However, in the colostrum from mothers who tested positive for IgG anti-G. lamblia, the level of this hormone had increased. The cortisol levels were similar between the groups, regardless of seropositivity. There was a higher percentage of Treg cells in the colostrum from mothers who tested positive for IgM anti-G. lamblia. TGF-β levels also increased in the colostrum of mothers who tested positive for IgM anti-G. lamblia. In the seronegative group for G. lamblia, there was a positive correlation between melatonin concentration and the percentage of Treg cells. These data suggest that the increase in regulatory cells and cytokines and the reduction in melatonin in colostrum from mothers with recent giardia infection may contribute to the evolution and manifestation of the disease. [ABSTRACT FROM AUTHOR]

Background: Giardiasis and zinc deficiency have been identified as serious health problems worldwide. Although Zn depletion is known to occur in giardiasis, no work has investigated whether changes occur in brain structures. Methods: Three groups of gerbils were used: control (1), orogastrically inoculated on day 3 after birth with trophozoites of two isolates of Giardia intestinalis (HGINV/WB) group (2 and 3). Estimates were made at five ages covering: establishment of infection, Giardia population growth, natural parasite clearance and a post-infection age. QuantiChrome zinc assay kit, cresyl violet staining and TUNEL technique were used. Results: A significant decrease (p<0.01) in tissue zinc was observed and persisted after infection. Cytoarchitectural changes were observed in 75% of gerbils in the HGINV or WB groups. Ectopic pyramidal neurons were found in the cornus ammonis (CA1-CA3). At 60 and 90 days of age loss of lamination was clearly visible in CA1. In the dentate gyrus (DG), thinning of the dorsal lamina and abnormal thickening of the ventral lamina were observed from 30 days of age. In the cerebellum, we found an increase (p<0.01) in the thickness of the external granular layer (EGL) at 14 days of age that persisted until day 21 (C 3 ± 0.3 μm; HGINV 37 ± 5 μm; WB 28 ± 3 μm); Purkinje cell population estimation showed a significant decrease; a large number of apoptotic somas were observed scattered in the molecular layer; in 60 and 90 days old gerbils we found granular cell heterotopia and Purkinje cell ectopia. The pattern of apoptosis was different in the cerebellum and hippocampus of parasitized gerbils. Conclusion: The morphological changes found suggest that neuronal migration is affected by zinc depletion caused by giardiasis in early postnatal life; for the first time, the link between giardiasis-zinc depletion and damaged brain structures is shown. This damage may explain the psychomotor/cognitive delay associated with giardiasis. These findings are alarming. Alterations in zinc metabolism and signalling are known to be involved in many brain disorders, including autism. Author summary: Giardia intestinalis (Syn. G. duodenalis or G. lamblia), is a protozoan flagellate that parasitizes humans and animals and is transmitted by the ingestion of food or water contaminated with cysts or via person-to-person contact. Causes diarrhoea. Affects children and adults worldwide. Giardiasis is known to have long-term effects. When giardiasis occurs in the first two years of a child's life, it is associated with stunted growth and psychomotor and cognitive retardation. Giardiasis and zinc deficiency have been identified as serious health problems worldwide. Although Zn depletion is known to occur in giardiasis, no work has investigated whether changes occur in brain structures. We modelled Giardia infection in gerbils in the first days of postnatal life and found morphological changes that suggest neuronal migration is affected by zinc depletion caused by giardiasis in early postnatal life. For the first time, the link between giardiasis-zinc depletion and damaged brain structures is shown. This damage may explain the psychomotor-cognitive delay associated with giardiasis. Alterations in zinc metabolism and signalling are known to be involved in many brain disorders, including autism. [ABSTRACT FROM AUTHOR]

A prospective study was conducted from September 2022 to March 2023 to examine the occurrence and contamination rates of different parasitic stages on fresh edible fruits and vegetables in local markets of Erbil city. Eight different types of fruits and vegetables were selected from eight markets, and 144 samples were analyzed for parasites using direct sedimentation, indirect floatation (ZnSO4), Kinyoun, and trichrome staining methods. The highest total frequencies of parasites were detected using the direct sedimentation method (18.75%), followed by the indirect floatation method (6.25%), Kinyoun technique (5.55%), and modified trichrome staining (4.16%). A. lumbricoides (Ova) was detected with 4.16% using direct sedimentation, while no positive detections were observed with other techniques. Kinyoun technique identified Cryptosporidium spp. oocysts. For E. histolytica 1.3% were positive using the direct sedimentation method, while the other three methods showed no positive detections. Similar results were observed for G. lamblia where only the modified trichrome staining technique detected 0.69% with trophozoites and 1.3% with cysts. Additionally, the direct sedimentation method revealed the presence of nematodes at 2.77%, but no positive detections were made with other methods. Concerning Taeniid eggs, the indirect floatation identified 0.69% with ova, while other methods showed no positive detections. Lastly, the direct sedimentation method yielded 6.25% with unidentified flagellate protozoa, but no positive detections were found using the indirect floatation method, Kinyoun technique, or modified trichrome staining technique. The study concluded that Kinyoun and trichrome techniques are the gold standard for detecting different parasite stages. [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]

Parasitic infections are a common problem in developing countries and can intensify morbidity in patients with sickle cell disease (SCD), increasing the severity of anemia and the need for transfusions. It has been demonstrated that both helminths and protozoa can affect gut microbiome composition. On the other hand, the presence of specific bacterial communities can also influence parasite establishment. Considering this, our aim was to associate the presence of intestinal parasites with the results of hematological analyses and microbiome composition evaluations in a population of Angolan children with and without SCD. A total of 113 stool samples were collected, and gut microbiome analysis was performed using 16S sequencing and real-time PCR to detect eight different intestinal parasites. In our population, more than half of children (55%) had at least one parasitic infection, and of these, 43% were co-infected. Giardia intestinalis and Ascaris lumbricoides were more frequently found in children from the rural area of Bengo. Moreover, SCD children with ascariasis exhibited higher values of leukocytes and neutrophils, whereas the total hemoglobin levels were lower. In regards to the gut microbiome, the presence of intestinal parasites lowered the prevalence of some beneficial bacteria, namely: Lactobacillus, Bifidobacterium, Cuneatibacter, Bacteroides uniformis, Roseburia, and Shuttleworthia. This study presents the prevalence of several intestinal parasites in a high-risk transmission area with scarce information and opens new perspectives for understanding the interaction between parasites, the microbiome, and SCD. [ABSTRACT FROM AUTHOR]

Gastrointestinal diseases of puppies aged 2-6 months account for 62% of all pathologies of these animals registered in the clinic. The lifestyle of dogs is associated with close tactile contact with the environment. The presence of protozoa in the digestive tract causes the inclusion of its representatives in the pathological process, which leads to the complication of disorders that have arisen due to various etiological factors. The study aimed to determine the role of the ubiquitous protozoa Cryptosporidium spp. in the pathogenesis of digestive tract diseases in puppies. General clinical, laboratory tests of blood (haematological, biochemical), faeces (coprogram, parasitological), statistical processing were performed. It was found that protozoa were detected in 52 % of sick animals with digestive disorders. The protozoan fauna included Cryptosporidium spp. (100 %), Isospora spp. (29 %), Giardia lamblia (21 %). Animals with cryptosporidium were examined without associations. The manifestation of the disease was noted most often in puppies with weak immune system functions, especially in the age group of 4-5 months, which is associated with the period of tooth change and a decrease in the overall immunological reactivity of the body. Clinically, the disease is characterized by enteritis development with accompanying symptoms. A characteristic clinical sign that should guide further diagnostic search is watery diarrhoea with an unpleasant odour, foamy, mucus, sometimes blood streaks. Laboratory findings include hypoalbuminemia with a decrease in the albumin-globulin ratio, hyperbilirubinaemia due to conjugated bilirubin, hyperfermentation of indicator enzymes of the liver and pancreas, and uremia. Such indicators indicate concomitant insufficiency of the digestive glands - the liver and pancreas, and a general inflammatory reaction. In coprological diagnostics, the qualitative data of the coprogramme confirm the lesions of the small intestine, liver and pancreas. Examination of fecal smears stained by the Ziehl-Nielsen method revealed cryptosporidium oocysts. The average intensity of invasion was 786.4 ± 23.1 oocysts per gram of feces. [ABSTRACT FROM AUTHOR]

Simple Summary: Blastocystis spp. and Giardia duodenalis are two prevalent intestinal parasites with a worldwide distribution that can infect humans and animals, resulting in significant public health concerns and economic losses. For goats, infection with the two parasites can cause symptoms such as diarrhea, which is not conducive to the development of stockbreeding. Shanxi Province is one of the largest goats breeding provinces in China. However, the prevalence of Blastocystis spp. and G. duodenalis in black goats in Shanxi Province remains unknown. Thus, 1200 fecal samples of black goats were collected in five representative geographical locations in Shanxi Province to examine the presence and genotypes of G. duodenalis and Blastocystis spp. by using a molecular approach. The results showed that the total infection rates of G. duodenalis and Blastocystis spp. were 7.5% and 3.5%, respectively. The presence and genotypes of G. duodenalis were determined based on three established loci (tpi, bg, and gdh). Among the detected assemblages B and E of G. duodenalis, the most prevalent assemblage was E in black goats in the five study areas. One novel MLG (MLG-E12) was identified by multilocus genotypes (MLGs) analysis. Through DNA sequence analysis, four subtypes of Blastocystis spp. were found in black goats, namely ST5, ST10, ST14, and ST30, among which ST10 was the dominant subtype in this study. This is the first report of Blastocystis spp. and G. duodenalis infection in black goats in Shanxi Province, which not only enhances our understanding of the genetic diversity of Blastocystis spp. and G. duodenalis in black goats in China but also provides essential baseline data for the prevention and control of Blastocystis spp. and G. duodenalis infection in black goats in the study areas. Blastocystis spp. and Giardia duodenalis are two prevalent zoonotic intestinal parasites that can cause severe diarrhea and intestinal diseases in humans and many animals. Black goat (Capra hircus) farming is increasingly important in China due to the remarkable adaptability, high reproductive performance, rapid growth rate, and significant economic value of black goats. A number of studies have indicated that black goats are the potential reservoir of multiple zoonotic protozoans in China; however, the prevalence and zoonotic status of G. duodenalis and Blastocystis spp. in black goats in Shanxi Province is still unknown. Thus, a total of 1200 fecal samples of black goats were collected from several representative regions at different altitudes in Shanxi Province and were examined for the presence and genotypes of G. duodenallis and Blastocystis spp. by amplifying the beta-giardin (bg), glutamate dehydrogenase (gdh), and triosephosphate isomerase (tpi) loci of G. duodenalis and SSU rRNA of Blastocystis spp. using PCR and sequence analysis methods, respectively. The overall prevalence of G. duodenalis and Blastocystis spp. in black goats in Shanxi Province were 7.5% and 3.5%, respectively. Two assemblages (B and E) of G. duodenalis and four subtypes (ST5, ST10, ST14, and ST30) of Blastocystis spp. were identified, with assemblage E and ST10 as the prevalent genotype and subtype in black goats, respectively. One novel multilocus genotype (MLG) was identified in MLG-E and was designated as MLG-E12. For both G. duodenalis and Blastocystis spp., the prevalence was significantly related to the region and age groups (p < 0.05). This is the first report on the prevalence of G. duodenalis and Blastocystis spp. in black goats in Shanxi Province. These results not only provide baseline data for the prevention and control of both parasites in black goats in Shanxi Province, but also enhance our understanding of the genetic composition and zoonotic potential of these two parasites. [ABSTRACT FROM AUTHOR]

Soil-transmitted helminths (STHs) are a group of parasites that are globally distributed and are the most prevalent neglected disease (NTD) in Latin America and the Caribbean (LAC); their presence is associated with health and development problems. In Argentina, their distribution is heterogenous, and there are highly endemic areas in the north of the country. The World Health Organization (WHO) recommends the mass deworming of children as a first-line strategy for the prevention and control of STHs and recently also encourage the development of more sensitive diagnostic tests. The aim of this study was to determine the prevalence of STHs in Tartagal (Salta, Argentina) after four years of deworming interruption with albendazole and ivermectin. A total of 437 fecal samples were analyzed using standard coprological techniques, a subset of which were selected to molecularly typify protozoan parasites; 257 blood samples were analyzed for the presence of specific antibodies to the STH Strongyloides stercoralis. The most prevalent protozoan species were G. intestinalis (19.6–49.2%) and B. hominis (19.1–38.5%). Molecular characterization allowed us to evidence possible zoonotic or human-to-human transmission pathways for Giardia intestinalis or Blastocystis spp., while serology for S. stercoralis proved to be a useful screening tool for monitoring this parasite after treatment. In general, a decrease in the prevalence of STHs was observed in the area, from 60% to 2.9–20% for hookworms and from 51% to 1–9.3% for S. stercoralis four years after treatment, demonstrating the effectiveness and duration of anthelmintic treatment with these two drugs. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

Parasitic diseases, predominantly prevalent in developing countries, are increasingly spreading to high-income nations due to shifting migration patterns. The World Health Organization (WHO) estimates approximately 300 million annual cases of giardiasis. The emergence of drug resistance and associated side effects necessitates urgent research to address this growing health concern. In this study, we evaluated over eleven thousand pharmacological compounds sourced from the FDA database to assess their impact on the TATA-binding protein (TBP) of the early diverging protist Giardia lamblia, which holds medical significance. We identified a selection of potential pharmacological compounds for combating this parasitic disease through in silico analysis, employing molecular modeling techniques such as homology modeling, molecular docking, and molecular dynamics simulations. Notably, our findings highlight compounds DB07352 and DB08399 as promising candidates for inhibiting the TBP of Giardia lamblia. Also, these compounds and DB15584 demonstrated high efficacy against trophozoites in vitro. In summary, this study identifies compounds with the potential to combat giardiasis, offering the prospect of specific therapies and providing a robust foundation for future research. [ABSTRACT FROM AUTHOR]

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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]

Background: At the end of 2022, there were over 108 million forcibly displaced people globally, including refugees, asylum seekers (AS) and internally displaced people (IDPs). Forced migration increases the risk of infectious disease transmission, and zoonotic pathogens account for 61% of emerging and re-emerging infectious diseases. Zoonoses create a high burden of disease and have the potential to cause large-scale outbreaks. This scoping review aimed to assess the state of research on a range of clinically relevant zoonotic pathogens in displaced populations in order to identify the gaps in literature and guide future research. Methodology / Principal findings: Literature was systematically searched to identify original research related to 40 selected zoonotic pathogens of interest in refugees, AS and IDPs. We included only peer-reviewed original research in English, with no publication date restrictions. Demographic data, migration pathways, health factors, associated outbreaks, predictive factors and preventative measures were extracted and synthesized. We identified 4,295 articles, of which 347 were included; dates of publications ranged from 1937 to 2022. Refugees were the most common population investigated (75%). Migration pathways of displaced populations increased over time towards a more complex web, involving migration in dual directions. The most frequent pathogen investigated was Schistosoma spp. (n = 99 articles). Disease outbreaks were reported in 46 publications (13.3%), with viruses being the most commonly reported pathogen type. Limited access to hygiene/sanitation, crowding and refugee status were the most commonly discussed predictors of infection. Vaccination/prophylaxis drug administration, surveillance/screening and improved hygiene/sanitation were the most commonly discussed preventative measures. Conclusions / Significance: The current research on zoonoses in displaced populations displays gaps in the spectrum of pathogens studied, as well as in the (sub)populations investigated. Future studies should be more inclusive of One Health approaches to adequately investigate the impact of zoonotic pathogens and identify transmission pathways as a basis for designing interventions for displaced populations. Author summary: Currently, there are over 108 million forcibly displaced people globally—mainly refugees, asylum seekers and internally displaced people. Migration is associated with increased exposure to infectious diseases. Zoonotic pathogens transmitted between humans and animals, account for the majority of emerging human diseases. Zoonoses are responsible for a substantial disease burden and are recognised to cause disease outbreaks, particularly in vulnerable populations in crowded living conditions. Unfortunately, zoonoses are often neglected by both healthcare services, medical research and especially in the context of migration. This review focussed on the current research of a spectrum of clinically relevant zoonotic pathogens in displaced populations to identify gaps in literature and inform on future research. Our findings highlighted the following gaps; zoonotic viruses appear particularly neglected, despite them being most commonly associated with disease outbreaks. The majority of publications investigating zoonoses in displaced people focused on three pathogens: Schistosoma species, Giardia lamblia and Leishmania species. There were very few publications investigating zoonotic pathogens in asylum seekers and internally displaced people compared to refugees. Future studies should be more inclusive and incorporate One Health approaches to adequately investigate the impact of zoonoses on displaced populations, and to identify transmission pathways for designing improved interventions. [ABSTRACT FROM AUTHOR]

Background: Intestinal parasitic infections (IP) are a major source of morbidity in people living with Human immunodeficiency virus (HIV), particularly in resource-limited settings, mostly as a result of high viral load. Hence, this study aimed to investigate the magnitude of intestinal parasitic infections and its determinants among patients with HIV/AIDS attending public health facilities in East and West Gojam Zones in Ethiopia. Methods: Institution-based cross-sectional study was conducted on 327 people living with HIV visiting public health facilities from December 2022 to May 2023. A simple random sampling technique was used to recruit participants. Face-to-face interviews were used to collect socio-demographics and determinants. The fresh stool was collected from each patient, transported, and tested in accordance with laboratory standard operating procedures of wet mount, formol-ether concentration technique, and modified acid-fast staining. Data were entered and analyzed in the statistical package for Social Science (SPSS) version 20. A 95% CI with p-value < 0.05 was considered statistically significant. Results: The overall prevalence of IP in patients with HIV/AIDS was 19.3% (63/327). Hookworm was the most identified parasite 33.3% (21/63) followed by E.histolytica 17% (11/63) and G.lamblia 14.3% (9/63). Parasitic infections were significantly higher among viral load > 1000cps/ml (p = 0.035), WHO stage 4 (p = 0.002), CD4 < 200 cell/mm3 (p = 0.001), and bare foot walking (p = 0.001). Conclusion: IP infections are moderately high among patients with HIV/AIDS in the study area. The proportion of parasites was greatly affected by high viral load, WHO stage 4, CD4 < 200 cell/mm3, and being barefoot; this gives valuable insight to health professionals, health planners and community health workers. As a result, viral load monitoring, and WHO stage controlling were periodically assessed in patients with HIV/AIDS. Health education, awareness creation, routine stool examination, and environmental hygiene were regularly advocated to increase the life of patients with HIV/AIDS. [ABSTRACT FROM AUTHOR]

Simple Summary: This research conducted in Serbia aimed to identify intestinal parasites in dogs that could potentially infect humans. Total prevalence of intestinal endoparasites was 62.6%. Various endoparasites such as Cystoisospora spp., Sarcocystis spp., Neospora caninum/Hammondia spp., Giardia intestinalis, Toxocara canis, Toxascaris leonina, Ancylostomatidae, Trichuris vulpis, Capillaria spp., Alaria alata and Taeniidae were found. Factors like age, outdoor living, attitude and diet were linked to higher infection rates. This study emphasizes the importance of educating dog owners, conducting routine parasitological tests on their pets and regular deworming strategies. Dogs are the most popular pets worldwide. Close contact between dogs and people increases the risk of transmission of various zoonotic parasitic infections. Given the importance of veterinary medicine in preserving the One Health concept, the aim of this research was to identify intestinal parasites that may have zoonotic potential and to evaluate risk factors (individual and environmental). The research was conducted in Serbia in 2022 and 2023 on 382 owned dogs, using qualitative methods of coprological examination with a concentration on parasitic elements. The overall prevalence of intestinal parasites was 62.6%, with the following detected: protozoa: Cystoisospora spp. (9.2%), Sarcocystis spp. (4.5%), Neospora caninum/Hammondia spp. (3.7%), Giardia intestinalis (11.8%); nematoda: Toxocara canis (11.5%), Toxascaris leonina (4.2%), family Ancylostomatidae (38.0%), Trichuris vulpis (21.5%), Capillaria spp. (10.5%); trematoda: Alaria alata (1.6%) and cestodes from the Taeniidae family (1.3%). Factors like age, size and coat length, as well as the way of living, attitude and diet were linked to a significantly higher (p < 0.05) prevalence of intestinal parasites. Based on the results of coprological diagnostics, this research indicates the importance of educating dog owners, conducting routine parasitological tests on their pets and regular deworming strategies. [ABSTRACT FROM AUTHOR]

An artificial semisynthetic material can be derived from artemisinin (ART) called dihydroartemisinin (DHA). Although DHA has enhanced antigiardial potential, its clinical application is limited because of its poor selectivity and low solubility. The drug's absorption has a direct impact on the cell, and mechanism research is limited to its destruction of the cytoskeleton. In this study, we used the zeolitic imidazolate framework-8 and loaded it with DHA (DHA@Zif-8) to improve its antigiardial potential. DHA@Zif-8 can enhance cellular uptake, increase antigiardial proliferation and encystation, and expand the endoplasmic reticulum compared with the DHA-treated group. We used RNA sequencing (RNA-seq) to investigate the antigiardial mechanism. We found that 126 genes were downregulated and 123 genes were upregulated. According to the KEGG and GO pathway analysis, the metabolic functions in G. lamblia are affected by DHA@Zif-8 NPs. We used real-time quantitative reverse transcription polymerase chain reaction to verify our results using the RNA-seq data. DHA@ Zif-8 NPs significantly enhanced the eradication of the parasite from the stool in vivo. In addition, the intestinal mucosal injury caused by G. lamblia trophozoites markedly improved in the intestine. This research provided the potential of utilizing DHA@Zif-8 to develop an antiprotozoan drug for clinical applications. [ABSTRACT FROM AUTHOR]

Background: Soil-transmitted helminthiasis, schistosomiasis and malaria remain a threat in developing countries, especially to people living in rural areas near large water bodies, valleys or swamps whose daily life activities revolve around those areas. In addition, these parasites affect different groups of people especially those in contact with their breeding sites including children mostly due to their developing immunity compared to that of old people. Methodology: This cross-sectional study was conducted in rural communities near the Rwasave and Cyarwa valleys, with 124 school-aged children participating. Stool samples were tested for the presence of STH eggs and intestinal Schistosoma parasites using the Kato Katz technique, and thick blood smears were made to aid in the identification of plasmodium species in blood. The threshold intensity of infection on STHs and Schistosoma sp. was estimated by counting their eggs microscopically, which aided in classifying infections as light, moderate, or heavy according to WHO standards. Furthermore, Plasmodium species were identified by the presence of schizonts, trophozoites, or gametocytes on Giemsa-stained thick blood smears, and parasitic density was calculated as a result. Results: In this study, the highest prevalence of soil transmitted helminthiasis was 12.7% in Cyarwa valley compared to 12.5% in Rwasave valley. The highest prevalence of Schistosomiasis was 3.2% in Cyarwa valley compared to 1.3% in Rwasave valley. Prevalence of malaria in Cyarwa valley was 3.3% in 63 children. Where children infected were only males falling in 5-8 age group. Conclusion: Results showed high prevalence of soil transmitted helminthiasis which was 12.7% in Cyarwa compared to 12.5% in Rwasave valley especially in children aged 5-8, this due to poor sanitation (P<0.001) that have assessed. Surprisingly, Cyarwa valley is located nearby town. Whereas malaria, intestinal schistosomiasis was low and there were not significantly associated with their risk factors with p-values 0.4, 0.092 respectively which can show that, preventive measures that have been established by government of Rwanda have positive impacts. Furthermore, there was a low prevalence of soil transmitted helminthiasis, intestinal schistosomiasis and malaria co-infection. [ABSTRACT FROM AUTHOR]