RESUMO
Entamoeba histolytica is a protozoan parasite belonging to the phylum Amoebozoa that causes amebiasis, a global public health problem. E. histolytica alternates its form between a proliferative trophozoite and a dormant cyst. Trophozoite proliferation is closely associated with amebiasis symptoms and pathogenesis whereas cysts transmit the disease. Drugs are available for clinical use; however, they have issues of adverse effects and dual targeting of disease symptoms and transmission remains to be improved. Development of new drugs is therefore urgently needed. An untargeted lipidomics analysis recently revealed structural uniqueness of the Entamoeba lipidome at different stages of the parasite's life cycle involving very long (26-30 carbons) and/or medium (8-12 carbons) acyl chains linked to glycerophospholipids and sphingolipids. Here, we investigated the physiology of this unique acyl chain diversity in Entamoeba, a non-photosynthetic protist. We characterized E. histolytica fatty acid elongases (EhFAEs), which are typically components of the fatty acid elongation cycle of photosynthetic protists and plants. An approach combining genetics and lipidomics revealed that EhFAEs are involved in the production of medium and very long acyl chains in E. histolytica. This approach also showed that the K3 group herbicides, flufenacet, cafenstrole, and fenoxasulfone, inhibited the production of very long acyl chains, thereby impairing Entamoeba trophozoite proliferation and cyst formation. Importantly, none of these three compounds showed toxicity to a human cell line; therefore, EhFAEs are reasonable targets for developing new anti-amebiasis drugs and these compounds are promising leads for such drugs. Interestingly, in the Amoebazoan lineage, gain and loss of the genes encoding two different types of fatty acid elongase have occurred during evolution, which may be relevant to parasite adaptation. Acyl chain diversity in lipids is therefore a unique and indispensable feature for parasitic adaptation of Entamoeba.
Assuntos
Entamoeba histolytica , Elongases de Ácidos Graxos , Elongases de Ácidos Graxos/metabolismo , Elongases de Ácidos Graxos/genética , Humanos , Entamoeba histolytica/efeitos dos fármacos , Entamoeba histolytica/genética , Proteínas de Protozoários/metabolismo , Proteínas de Protozoários/genética , Entamoeba/efeitos dos fármacos , Entamoeba/metabolismo , Amebíase/tratamento farmacológico , Amebíase/parasitologia , Entamebíase/parasitologia , Entamebíase/tratamento farmacológico , Entamebíase/metabolismo , Trofozoítos/efeitos dos fármacos , Trofozoítos/metabolismo , Antiprotozoários/farmacologia , Ácidos Graxos/metabolismoRESUMO
Entamoeba moshkovskii, according to recent studies, appears to exert a more significant impact on diarrhoeal infections than previously believed. The efficient identification and genetic characterization of E. moshkovskii isolates from endemic areas worldwide are crucial for understanding the impact of parasite genomes on amoebic infections. In this study, we employed a multilocus sequence typing system to characterize E. moshkovskii isolates, with the aim of assessing the role of genetic variation in the pathogenic potential of E. moshkovskii. We incorporated 3 potential genetic markers: KERP1, a protein rich in lysine and glutamic acid; amoebapore C (apc) and chitinase. Sequencing was attempted for all target loci in 68 positive E. moshkovskii samples, and successfully sequenced a total of 33 samples for all 3 loci. The analysis revealed 17 distinct genotypes, labelled M1M17, across the tested samples when combining all loci. Notably, genotype M1 demonstrated a statistically significant association with diarrhoeal incidence within E. moshkovskii infection (P = 0.0394). This suggests that M1 may represent a pathogenic strain with the highest potential for causing diarrhoeal symptoms. Additionally, we have identified a few single-nucleotide polymorphisms in the studied loci that can be utilized as genetic markers for recognizing the most potentially pathogenic E. moshkovskii isolates. In our genetic diversity study, the apc locus demonstrated the highest Hd value and π value, indicating its pivotal role in reflecting the evolutionary history and adaptation of the E. moshkovskii population. Furthermore, analyses of linkage disequilibrium and recombination within the E. moshkovskii population suggested that the apc locus could play a crucial role in determining the virulence of E. moshkovskii.
Assuntos
Entamoeba , Tipagem de Sequências Multilocus , Marcadores Genéticos , Entamoeba/genética , Entamoeba/classificação , Entamoeba/isolamento & purificação , Humanos , Entamebíase/parasitologia , Entamebíase/epidemiologia , Genótipo , Polimorfismo de Nucleotídeo Único , Variação Genética , FilogeniaRESUMO
BACKGROUND: Parasites Entamoeba spp., Enterocytozoon bieneusi and Blastocystis are prevalent pathogens causing gastrointestinal illnesses in animals and humans. Consequently, researches on their occurrence, distribution and hosts are crucial for the well-being of both animals and humans. Due to the confined spaces and frequent interaction between animals and humans, animal sanctuaries have emerged as potential reservoirs for these parasites. In this study, the wildlife sanctuary near the Huang Gorge of the Qinling Mountains in northwest China is chosen as an ideal site for parasite distribution research, considering its expansive stocking area and high biodiversity. RESULTS: We collected 191 fecal specimens from 37 distinct wildlife species and extracted genomic DNA. We identified these three parasites by amplifying specific gene regions and analyzed their characteristics and evolutionary relationships. All the parasites exhibited a high overall infection rate, reaching 90.05%. Among them, seven Entamoeba species were identified, accounting for a prevalence of 54.97%, with the highest infection observed in Entamoeba bovis. In total, 11 Enterocytozoon bieneusi genotypes were discovered, representing a prevalence of 35.08%, including three genotypes of human-pathogenic Group 1 and two novel genotypes (SXWZ and SXLG). Additionally, 13 Blastocystis subtypes were detected, showing a prevalence of 74.87% and encompassing eight zoonotic subtypes. All of the above suggests significant possibilities of parasite transmission between animals and humans. CONCLUSIONS: This study investigated the occurrence and prevalence of three intestinal parasites, enhancing our understanding of their genetic diversity and host ranges in northwest China. Furthermore, the distribution of these parasites implies significant potential of zoonotic transmission, underscoring the imperative for ongoing surveillance and implementation of control measures. These efforts are essential to mitigate the risk of zoonotic disease outbreaks originating from wildlife sanctuary.
Assuntos
Animais Selvagens , Blastocystis , Entamoeba , Enterocytozoon , Microsporidiose , Zoonoses , Animais , Enterocytozoon/genética , Enterocytozoon/isolamento & purificação , China/epidemiologia , Blastocystis/genética , Blastocystis/classificação , Blastocystis/isolamento & purificação , Animais Selvagens/parasitologia , Zoonoses/parasitologia , Entamoeba/genética , Entamoeba/isolamento & purificação , Entamoeba/classificação , Microsporidiose/veterinária , Microsporidiose/epidemiologia , Filogenia , Fezes/parasitologia , Entamebíase/veterinária , Entamebíase/epidemiologia , Entamebíase/parasitologia , Infecções por Blastocystis/veterinária , Infecções por Blastocystis/epidemiologia , Infecções por Blastocystis/transmissão , Infecções por Blastocystis/parasitologia , Prevalência , Genótipo , HumanosRESUMO
Amoebiasis is a disease caused by Entamoeba histolytica, affecting the large intestine of humans and occasionally leading to extra-intestinal lesions. Entamoeba dispar is another amoeba species considered commensal, although it has been identified in patients presenting with dysenteric and nondysenteric colitis, as well as amoebic liver abscess. Amoebic virulence factors are essential for the invasion and development of lesions. There is evidence showing that the association of enterobacteria with trophozoites contributes to increased gene expression of amoebic virulence factors. Enteropathogenic Escherichia coli is an important bacterium causing diarrhea, with high incidence rates in the world population, allowing it to interact with Entamoeba sp. in the same host. In this context, this study aims to evaluate the influence of enteropathogenic Escherichia coli on ACFN and ADO Entamoeba dispar strains by quantifying the gene expression of virulence factors, including galactose/N-acetyl-D-galactosamine-binding lectin, cysteine proteinase 2, and amoebapores A and C. Additionally, the study assesses the progression and morphological aspect of amoebic liver abscess and the profile of inflammatory cells. Our results demonstrated that the interaction between EPEC and ACFN Entamoeba dispar strains was able to increase the gene expression of virulence factors, as well as the lesion area and the activity of the inflammatory infiltrate. However, the association with the ADO strain did not influence the gene expression of virulence factors. Together, our findings indicate that the interaction between EPEC, ACFN, and ADO Entamoeba dispar strains resulted in differences in vitro and in vivo gene expression of Gal/GalNAc-binding lectin and CP2, in enzymatic activities of MPO, NAG, and EPO, and consequently, in the ability to cause lesions.
Assuntos
Entamoeba , Escherichia coli Enteropatogênica , Fatores de Virulência , Escherichia coli Enteropatogênica/patogenicidade , Escherichia coli Enteropatogênica/genética , Entamoeba/patogenicidade , Entamoeba/genética , Entamoeba/fisiologia , Fatores de Virulência/genética , Virulência , Animais , Camundongos , Abscesso Hepático Amebiano/parasitologia , Entamebíase/parasitologia , Humanos , Expressão GênicaRESUMO
Vegetable and fruit contamination is recognized as a significant parasite transmission route. This review presents the current state of vegetables ad fruits contamination with food-borne parasitic protozoa worldwide. We consider the methodologies and strategies for detecting parasitic stages developed in the last decade and the contamination data. Asia had the highest number of reports (94 studies), followed by Africa (74 studies). At the country level, with 41 studies, Iran had the most reports among other countries, followed by Nigeria (28 studies). According to the studies included in the current review, 41.22% of vegetables and fruits were contaminated with different species of protozoan parasites. Among different continents, Asia accounted for the highest contamination rate of protozoan parasites (57.12%). Giardia spp. (10%) had the highest contamination rate in vegetables and fruits, followed by Entamoeba coli (8%), E. histolytica/dispar (7%), and Cryptosporidium spp. (6%). This study provides essential data for health authorities to develop food safety programs. The presence of protozoan parasites in fruits and vegetables highlights the critical need for maintaining rigorous food safety measures across the entire production and distribution process, particularly in countries that are major producers and distributors of these food items.
Assuntos
Contaminação de Alimentos , Frutas , Verduras , Verduras/parasitologia , Frutas/parasitologia , Contaminação de Alimentos/análise , Humanos , Animais , Inocuidade dos Alimentos , Parasitologia de Alimentos , Cryptosporidium/isolamento & purificação , Cryptosporidium/genética , Parasitos/isolamento & purificação , Parasitos/classificação , Parasitos/genética , Giardia/isolamento & purificação , Giardia/genética , Entamoeba/isolamento & purificação , Entamoeba/genética , ÁsiaRESUMO
The parasite Entamoeba histolytica is the cause of amoebic dysentery and liver abscess in humans. On the protozoan cell surface, a variety of glycosylated molecules are involved in the interaction with the environment, such as attachment to the colonic mucus. One of these molecules is the lipopeptidophosphoglycan (LPPG), a complex surface component with antigenic properties. Its structure is only partly known, it is a glycosylphosphatidylinositol (GPI)-linked glycoprotein with a large amount of O-glycosylation. To date, the sequence of a core protein has not been identified. In this study, we further investigated this complex surface molecule aided by the availability of the monoclonal antibody EH5, which had been raised in our laboratory. We studied the extraction of LPPG in various solvent mixtures and discovered that 2-butanol saturated water was simple and superior to other solvents used in the past. The isolated LPPG was subjected to treatment with several proteases and the Ser/Thr specific cleavage agent scandium (III) trifluoromethanesulfonate (scandium triflate). The products were probed with antibody EH5 and the blots showed that the LPPG preparation was largely resistant to standard proteases, but could be cleaved by the scandium compound. These observations could point to the existence of a Ser- or Thr-rich core protein structure.
Assuntos
Entamoeba histolytica , Entamoeba , Peptidoglicano , Fosfolipídeos , Humanos , Escândio , Antígenos de Protozoários , Peptídeo HidrolasesRESUMO
The oral cavity is a habitat to a diverse range of organisms that make up an essential element of the human microbiota. There are up to 1000 species of micro-organisms capable of colonizing the mouth. Thirty percent of them are uncultivable. The genus Entamoeba includes several species, out of which at least seven of them are able to inhabit the human body (Entamoeba histolytica, Entamoeba dispar, Entamoeba moshkovskii, Entamoeba coli, Entamoeba polecki, Entamoeba hartmann, Entamoeba gingivalis). It was shown that only E. gingivalis is able to colonize the oral cavity. The aim of this study was to evaluate the association and prevalence of E. gingivalis in periodontal disease using two electronic database search engines. In order to have a broader view of the subject, a comprehensive manual search was conducted between 15th February 2023 and 1 April 2023 on these content aggregators and the initial search resulted in 277 articles using the keywords "E. gingivalis", "periodontitis", "E. gingivalis", "periodontal disease", "prevalence", and "incidence", in different combinations. The results showed that 755 patients were infected with E. gingivalis out of a total number of 1729 patients diagnosed with periodontal disease, indicating a global prevalence of 43% in the set of patients analyzed. E. gingivalis was prevalent in 58% of the patients that had gingivitis and in 44% of the patients with periodontitis. Prevalence of E. gingivalis based on gender was 43% in female patients and 47% in male patients. The results indicate that the higher incidence of E. gingivalis in people with periodontal disease compared to healthy people is more than just a sign of the disease; it could also be linked to the severity of the condition and the disease propensity to progress.
Assuntos
Entamoeba , Doenças Periodontais , Humanos , Entamoeba/isolamento & purificação , Entamoeba/patogenicidade , Doenças Periodontais/microbiologia , Doenças Periodontais/epidemiologia , Entamebíase/epidemiologia , Prevalência , Feminino , MasculinoRESUMO
Amoebic dysentery is a common infectious disease that is acquired through contaminated food and water harboring the infective stage of the parasite. Entamoeba histolytica is a parasite that is spread globally causing increasing morbidity and mortality in developing countries. To Identify the infection of Entamoeba histolytica by PCR and other lab methods among patients attending Kirkuk hospitals. Methods: The current study involved the examination of 220 fecal specimens from children under 15 years during the period of 1st January 2023 until 5th of June 2023. It involved microscopic examination of fecal samples confirmation of diagnosis with two different ELISA tests that capture E. histolytic. Also, microscopic positive samples were submitted to nucleic acid detection of E. histolytic via Real-Time PCR. The percentage of positive specimens that were tested with E. histolytica / dispar ELISA (DRG ELISA), out of 93 stool specimens, 59 (63.44%) were positive, while the remaining specimens 34 (36.56%) were negative despite being tested positive by microscopy. The DRG stool ELISA revealed sensitivity and specificity (69.28% and 97.91%) respectively and a predictive value of (97%). The sample that was the positive result with DRG ELISA was discriminated via Tech Lab E. histolytica ELISA which detects the presence of only E. histolytica alone in fecal samples. Out of 93 examined specimens, only 24 (25.81%) were positive while the remaining 69 (74.19%) were negative. DRG ELISA for E. histolytica/dispar positive results were 63.44%, while TechLab ELISA has produced 25.81% positive E. histolytica. Whereases, RT PCR results were only 20.44%. Qi square analysis was applied and yielded a significant difference v=between the method of diagnosis with P=<0.0001. Microscopy-positive Entamoeba complex is a primitive means of detection of Entamoeba complex and diagnosis should always be confirmed with superior method like ELISA or PCR.
Assuntos
Entamoeba histolytica , Entamoeba , Entamebíase , Humanos , Criança , Entamebíase/diagnóstico , Entamebíase/parasitologia , Entamoeba histolytica/genética , Sensibilidade e Especificidade , Reação em Cadeia da Polimerase em Tempo Real , Ensaio de Imunoadsorção Enzimática , Fezes/parasitologia , Diarreia/diagnósticoRESUMO
The protozoan parasite Entamoeba histolytica causes amoebiasis, a global public health problem. Amoebiasis is solely transmitted by cysts that are produced from proliferative trophozoites by encystation in the large intestine of humans. During encystation, various metabolites, pathways, and cascades sequentially orchestrate the morphological and physiological changes required to produce cysts. Cholesteryl sulfate (CS) has recently been revealed to be among the key molecules that control the morphological and physiological changes of encystation by exerting pleiotropic effects. CS promotes the rounding of encysting Entamoeba cells and maintains this spherical morphology as encysting cells are surrounded by the cyst wall, a prerequisite for resistance against environmental stresses. CS is also involved in the development of membrane impermeability, another prerequisite for resistance. The initiation of cyst wall formation is, however, CS-independent. Here, we overview CS-dependent and -independent processes during encystation and discuss their functional linkage. We also discuss a potential transcriptional cascade that controls the processes necessary to produce dormant Entamoeba cysts.
Assuntos
Amebíase , Cistos , Entamoeba histolytica , Entamoeba , Humanos , Entamoeba/metabolismoRESUMO
Entamoeba histolytica, the causative agent of amebiasis, is the third leading cause of death among parasitic diseases globally. Its life cycle includes encystation, which has been mostly studied in Entamoeba invadens, responsible for reptilian amebiasis. However, the molecular mechanisms underlying this process are not fully understood. Therefore, we focused on the identification and characterization of Myb proteins, which regulate the expression of encystation-related genes in various protozoan parasites. Through bioinformatic analysis, we identified 48 genes in E. invadens encoding MYB-domain-containing proteins. These were classified into single-repeat 1R (20), 2R-MYB proteins (27), and one 4R-MYB protein. The in-silico analysis suggests that these proteins are multifunctional, participating in transcriptional regulation, chromatin remodeling, telomere maintenance, and splicing. Transcriptomic data analysis revealed expression signatures of eimyb genes, suggesting a potential orchestration in the regulation of early and late encystation-excystation genes. Furthermore, we identified probable target genes associated with reproduction, the meiotic cell cycle, ubiquitin-dependent protein catabolism, and endosomal transport. In conclusion, our findings suggest that E. invadens Myb proteins regulate stage-specific proteins and a wide array of cellular processes. This study provides a foundation for further exploration of the molecular mechanisms governing encystation and unveils potential targets for therapeutic intervention in amebiasis.
Assuntos
Amebíase , Entamoeba histolytica , Entamoeba , Humanos , Entamoeba/genética , Entamoeba/metabolismo , Entamoeba histolytica/genética , Perfilação da Expressão Gênica , Regulação da Expressão GênicaRESUMO
Entamoeba histolytica infects the large intestine of humans, causing a spectrum of clinical appearances ranging from asymptomatic colonization to severe intestinal and extra-intestinal disease. The parasite is identical microscopically to commensal nonpathogenic amoeba. To detect the pathogenic Entamoeba and estimate the precise prevalence of the parasite among the symptomatic pediatric population using molecular techniques. 323 fecal samples were collected from symptomatic children admitted to Sulaimani Pediatric Teaching Hospital, Sulaimaniyah Province, Iraq, from June to October 2021. A structured, validated questionnaire was prepared and used to report participants' gender, residency, and drinking water source. Then, stool samples were microscopically examined, and the positive samples were submitted to molecular analysis by amplifying the 18s rRNA gene using nested PCR to differentiate E. histolytica from other nonpathogenic Entamoeba. Finally, gene sequences were done to confirm the species. Microscopically, 58 positive samples represented Entamoeba species infection rate of 18% among symptomatic patients. However, only 18 samples were positive for E. histolytica based on molecular methods, which accounts for 31% of the positive by microscopy and 5.6% among the 323 symptomatic populations. NCBI, available in their database, gives the gene sequence and accession number. Patients' sociodemographic data and water sources were directly related to the infection rate. Classical microscopic examination provides a misleading profile about the prevalence of E. histolytica in an endemic region that might lead to unnecessary treatments and a lack of appropriate management for patients.
Assuntos
Entamoeba , Entamebíase , Humanos , Criança , Entamoeba/genética , Iraque/epidemiologia , Entamebíase/epidemiologia , Fezes , HospitalizaçãoRESUMO
Wild rhesus macaques are a potential source of zoonotic parasites for humans, and Entamoeba spp. are common intestinal parasites. To investigate the prevalence of Entamoeba in wild rhesus macaques in China and explore the genetic differentiation of the potentially pathogenic species Entamoeba nuttalli, a total of 276 fecal samples from five populations at high altitudes (HAG, 2,800-4,100 m above sea level) and four populations at low altitudes (LAG, 5-1,000 m above sea level) were collected. PCR methods based on the ssrRNA gene were used to detect Entamoeba infection. Genotyping of E. nuttalli was performed based on six tRNA-linked short tandem repeat (STR) loci for further genetic analyses. The results revealed that Entamoeba infection (69.2%) was common in wild rhesus macaques in China, especially in LAG which had a significantly higher prevalence rate than that in HAG (P < 0.001). Three zoonotic species were identified: Entamoeba chattoni (60.9%) was the most prevalent species and distributed in all the populations, followed by Entamoeba coli (33.3%) and Entamoeba nuttalli (17.4%). In addition, a novel Entamoeba ribosomal lineage named RL13 (22.8%) was identified, and phylogenetic analysis revealed a close genetic relationship between RL13 and Entamoeba. hartmanni. Genotyping of E. nuttalli obtained 24 genotypes from five populations and further analysis showed E. nuttalli had a high degree of genetic differentiation (FST > 0.25, Nm < 1) between the host populations. The result of analysis of molecular variance (AMOVA) revealed that observed genetic differences mainly originate from differences among populations (FST = 0.91). Meanwhile, the phylogenetic tree showed that these genotypes of E. nuttalli were clustered according to geographical populations, indicating a significant phylogeographic distribution pattern. Considering the potential pathogenicity of E. nuttalli, attention should be paid to its risk of zoonotic transmission.
Assuntos
Entamoeba , Entamebíase , Fezes , Genótipo , Macaca mulatta , Filogenia , Animais , Entamoeba/genética , Entamoeba/classificação , Entamoeba/isolamento & purificação , China/epidemiologia , Entamebíase/epidemiologia , Entamebíase/parasitologia , Entamebíase/veterinária , Fezes/parasitologia , Doenças dos Macacos/parasitologia , Doenças dos Macacos/epidemiologia , Prevalência , Variação Genética , Repetições de Microssatélites , DNA de Protozoário/genéticaRESUMO
The study involved the estimation of the prevalence of Entamoeba spp. using microscopy and molecular techniques among symptomatic outpatients during April 2021 to March, 2022. Stool samples were collected from 2592 outpatients with amoebiasis symptoms of both sexes and different ages (≤ l to 60). Also, 107 stool samples were taken randomly from asymptomatic individuals and examined microscopically to detect infection with Entamoeba spp. the positive specimens were used for molecular analysis with positive symptomatic samples targeting the 18S rRNA gene by nested PCR. Microscopically 21.68% (562/2592) were positive, for Entamoeba spp. Males showed highest infection rate than females (67.43% vs 32.56%). Ages from 1-10 years showed the highest rate (54.09%), and urban inhabitant had somewhat a higher rate than rural one (58.54% vs 41.45%) which was statistically non-significant(P>0.05). Among asymptomatic individuals, 57% (61/107) were positive for Entamoeba spp. Nested PCR analysis yielded 73% positive samples for Entamoeba spp. with a fragment size of 897 bp. Three fragment sizes were produced, for E. histolytica, E. dispar and E. moshkovskii which were 439, 174 and 553 bps, respectively. Single infection occurred with, E. histolytica in 46%, of symptomatic and 6% of asymptomatic cases, E. dispar in 38% of asymptomatic and 10% of symptomatic cases, E. moshkovskii, reported at very low rate among both groups.
Assuntos
Entamoeba , Entamebíase , Fezes , Humanos , Fezes/parasitologia , Iraque/epidemiologia , Criança , Pré-Escolar , Masculino , Entamoeba/isolamento & purificação , Entamoeba/genética , Entamoeba/classificação , Feminino , Adolescente , Adulto , Lactente , Pessoa de Meia-Idade , Adulto Jovem , Entamebíase/epidemiologia , Entamebíase/parasitologia , Entamebíase/diagnóstico , Reação em Cadeia da Polimerase , RNA Ribossômico 18S/genética , Prevalência , DNA de Protozoário/genéticaRESUMO
In humans, Entamoeba histolytica is the main pathogen causing various amoebiases, while E. moshkovskii falls between being a pathogen and non-pathogen. The two species have similar behavior patterns but differ significantly in pathogenicity, with previous studies and clinical data indicating that E. moshkovskii has a low level of pathogenicity. Meaningfully, the biological characteristics of E. moshkovskii make it a potential model organism and a protein display platform for studying the functions of important Entamoeba proteins. Here, an Amoeba-pcDNA3.1 vector capable of overexpressing E. histolytica-sourced Igl-C protein was constructed and successfully transfected into E. moshkovskii. High levels of expression of the Igl-C, EGFP, and NeoR genes were identified in Igl-C-transfected trophozoites using qRT-PCR, and they were subsequently confirmed using immunoblotting. Transfection of Igl-C protein improved the adherence and phagocytosis of E. moshkovskii, demonstrating that E. histolytica Igl mediated amoebic adhesion. Moreover, as a manifestation of protein virulence, the ability of post-transfected trophozoites to induce inflammation in host macrophages was also enhanced. In conclusion, this study utilizing the characteristics of E. moshkovskii confirmed its potential to serve as a model organism. E. moshkovskii could replace E. histolytica as the target of gene editing, allowing more efficient study of amoebic pathogenicity.
Assuntos
Entamoeba histolytica , Entamoeba , Proteínas de Protozoários , Trofozoítos , Entamoeba/genética , Entamoeba/patogenicidade , Proteínas de Protozoários/genética , Proteínas de Protozoários/metabolismo , Entamoeba histolytica/genética , Entamoeba histolytica/patogenicidade , Entamoeba histolytica/metabolismo , Trofozoítos/metabolismo , Fagocitose , Lectinas/metabolismo , Lectinas/genética , Humanos , Animais , Transfecção , Virulência/genética , Entamebíase/parasitologia , CamundongosRESUMO
Entamoeba moshkovskii, recently known as a possible pathogenic amoeba, and the non-pathogenic Entamoeba dispar are morphologically indistinguishable by microscopy. Although PCR was used for differential diagnosis, gel electrophoresis is labor-intensive, time-consuming, and exposed to hazardous elements. In this study, nucleic acid lateral flow immunoassay (NALFIA) was developed to detect E. moshkovskii and E. dispar by post-PCR amplicon analysis. E. moshkovskii primers were labeled with digoxigenin and biotin whereas primers of E. dispar were lebeled with FITC and digoxigenin. The gold nanoparticles were labeled with antibodies corresponding to particular labeling. Based on the established assay, NALFIA could detect as low as 975 fg of E. moshkovskii target DNA (982 parasites or 196 parasites/microliter), and 487.5 fg of E. dispar target DNA (444 parasites or 89 parasites/microliter) without cross-reactivity to other tested intestinal organisms. After testing 91 stool samples, NALFIA was able to detect seven E. moshkovskii (87.5% sensitivity and 100% specificity) and eight E. dispar samples (66.7% sensitivity and 100% specificity) compared to real-time PCR. Interestingly, it detected three mixed infections as real-time PCR. Therefore, it can be a rapid, safe, and effective method for the detection of the emerging pathogens E. moshkovskii and E. dispar in stool samples.
Assuntos
Amoeba , Entamoeba histolytica , Entamoeba , Entamebíase , Nanopartículas Metálicas , Ácidos Nucleicos , Humanos , Entamoeba/genética , Entamebíase/diagnóstico , Entamebíase/parasitologia , Amoeba/genética , Digoxigenina , Ouro , DNA de Protozoário/genética , DNA de Protozoário/análise , Reação em Cadeia da Polimerase em Tempo Real , Imunoensaio , Fezes/química , Entamoeba histolytica/genéticaRESUMO
Amoebiasis, caused by the enteric parasite, Entamoeba histolytica, is one of the major food- and water-borne parasitic diseases in developing countries with improper sanitation and poor hygiene. Infection with E. histolytica has diverse disease outcomes, which are determined by the genetic diversity of the infecting strains. Comparative genetic analysis of infecting E. histolytica strains associated with differential disease outcomes from different geographical regions of the world is important to identify the specific genetic patterns of the pathogen that trigger certain disease outcomes of Amoebiasis. The strategy is able to elucidate the genealogical relation and population structure of infecting E. histolytica strains from different geographical regions. In the present study, we have performed a comparative genetic analysis of circulating E. histolytica strains identified from different parts of the world, including our study region, based on five tRNA-linked short tandem repeat (STR) loci (i.e., D-A, NK2, R-R, STGA-D and A-L) and evaluated their potential associations with differential disease outcomes of Amoebiasis. A number of regional-specific, emerging haplotypes of E. histolytica, significantly associated with specific disease outcomes have been identified. Haplotypes, which have a significant positive association with asymptomatic and amoebic liver abscess outcomes, showed a significant negative association with diarrheal outcome, or vice versa. Comparative multi-locus analysis revealed that E. histolytica isolates from our study region are phylogenetically segregated from the isolates of other geographical regions. This study provides a crucial overview of the population structure and emerging pattern of the enteric parasite, E. histolytica.
Assuntos
Amebíase , Disenteria Amebiana , Entamoeba histolytica , Entamoeba , Entamebíase , Abscesso Hepático Amebiano , Animais , Entamoeba histolytica/genética , Entamebíase/epidemiologia , Entamebíase/parasitologia , Abscesso Hepático Amebiano/parasitologia , Disenteria Amebiana/parasitologia , Análise de Sequência , Entamoeba/genéticaRESUMO
Amoebiasis, caused by the enteric parasite Entamoeba histolytica has differential disease outcomes. The association of parasite genotypes with outcomes of amoebic infection is still a paradox and requires to be explored. The genetic information of infecting strains from endemic settings of different geographical regions is essential to evaluate the relation. Comparative genetics of E. histolytica clinical isolates from different disease outcomes have been explored based on two tRNA-linked STR loci (STGA-D and A-L). All of the repeat patterns in the A-L locus were newly identified and unique to Indian isolates. The majority of newly identified repeat patterns in STGA-D locus have outcome-specific distributions, predicting the emergence of disease-specific mutations in this target locus. Statistical analysis further reinforces this observation, as identified repeat patterns only from STGA-D but not A-L locus were significantly associated with disease outcomes. Phylogenetic analysis indicates independent segregation and divergence of tRNA-linked STR arrays for each STR locus.
Assuntos
Entamoeba histolytica , Entamoeba , Entamebíase , Animais , Entamoeba histolytica/genética , Entamebíase/epidemiologia , Entamebíase/parasitologia , Marcadores Genéticos , Filogenia , Repetições de Microssatélites , RNA de Transferência/genética , Entamoeba/genéticaRESUMO
Despite great efforts, intestinal protozoan infections remain a significant healthcare concern worldwide. Although many point-of-care (POC) tests are increasingly being used, microscopic examination of stool specimens remains the mainstay for their diagnosis, especially in resource-limited settings. We assessed the utility of rapid POC tests based on immunochromatography among patients from rural Northern India. A total of 78 patients were enrolled in the study. Out of nine specimens that tested positive for Giardia duodenalis on microscopy, an immunochromatographic test (ICT) could detect only five (55.55%). Entamoeba histolytica/dispar was demonstrated in two specimens on microscopy, both of which were missed by ICT. Its overall sensitivity, specificity, and positive and negative predictive value were 50%, 98.5%, 83.3%, and 93%, respectively. Its performance was considered unsatisfactory. Although ICT-based tests provide a relatively rapid and less labor-intensive alternative, they should be used to supplement and not replace stool microscopy.
Assuntos
Entamoeba histolytica , Entamoeba , Entamebíase , Infecções por Protozoários , Humanos , Pacientes Ambulatoriais , Entamebíase/diagnóstico , Entamebíase/epidemiologia , Fezes , Sensibilidade e Especificidade , Infecções por Protozoários/diagnósticoRESUMO
Despite extensive research, little is known about the composition of eukaryotic protists in environmental samples. This is due to low parasite concentrations, the complexity of parasite diversity, and a lack of suitable reference databases and standardized protocols. To bridge this knowledge gap, this study used 18S rRNA short amplicon and shotgun metagenomic sequencing approaches to profile protozoan microbial communities as well as their functional pathways in treated and untreated wastewater samples collected from different regions of South Africa. Results demonstrated that protozoan diversity (Shannon index P-value = 0.03) and taxonomic composition (PERMANOVA, P-value = 0.02) was mainly driven by the type of wastewater samples (treated & untreated) and geographic location. However, these WWTPs were also found to contain a core community of protozoan parasites. The untreated wastewater samples revealed a predominant presence of free-living, parasitic, and potentially pathogenic protists typically found in humans and animals, ranging from Alveolata (27 %) phylum (Apicomplexa and Ciliophora) to Excavata (3.88 %) (Discoba and Parasalia) and Amoebozoa (2.84 %) (Entamoeba and Acanthamoeba). Shotgun metagenomics analyses in a subset of the untreated wastewater samples confirmed the presence of public health-importance protozoa, including Cryptosporidium species (3.48 %), Entamoeba hystolitica (6.58 %), Blastocystis hominis (2.91 %), Naegleria gruberi (2.37 %), Toxoplasma gondii (1.98 %), Cyclospora cayetanensis (1.30 %), and Giardia intestinalis (0.31 %). Virulent gene families linked to pathogenic protozoa, such as serine/threonine protein phosphatase and mucin-desulfating sulfatase were identified. Additionally, enriched pathways included thiamine diphosphate biosynthesis III, heme biosynthesis, Methylerythritol 4-Phosphate Pathway, methyl erythritol phosphate (MEP), and pentose phosphate pathways. These findings suggest that protozoan pathogens may possess metabolic and growth potential within WWTPs, posing a severe risk of transmission to humans and animals if inadequately disinfected before release. This study provides a baseline for the future investigation of diverse protozoal communities in wastewater, which are of public health importance.
Assuntos
Criptosporidiose , Cryptosporidium , Entamoeba , Parasitos , Animais , Humanos , Águas Residuárias , Cryptosporidium/genética , RNA Ribossômico 18S , Eucariotos , Metagenômica/métodos , Fosfatos/análiseRESUMO
Introduction: Children with intellectual disability (ID) often face challenges in maintaining proper oral hygiene due to their motor, sensory, and intellectual impairments, which can lead to compromised oral health; therefore, there is a need to enhance the oral health status of these populations and establish an effective system for administering preventive interventions. Here, we aimed to evaluate the prevalence of Entamoeba gingivalis and Trichomonas tenax among children with ID in Lorestan province, in Western Iran through parasitological and molecular methods. Methods: The current descriptive investigation involved 215 in children with ID and 215 healthy children (non-ID) who were referred to health facilities in Lorestan province, Iran between October 2022 and March 2024. The prevalence of protozoa in the oral cavity was found through the utilization of both microscopic analysis and conventional polymerase chain reaction (PCR) techniques. Results: The total prevalence of the E. gingivalis and T. tenax in children with ID was found to be 87 (40.5%) and 92 (42.8%) through microscopic and PCR methods, respectively. Among the positive samples, 57 (61.9%) and 35 (38.1%) children tested positive for E. gingivalis and T. tenax, respectively. In contrast, among the 215 non-ID children in the control group, 39 (18.1%) and 42 (19.5%) tested positive by microscopic and PCR methods, respectively. Among positive samples in non-ID children, 23 (54.7%) and 19 (45.3%) children were positive for E. gingivalis and T. tenax, respectively. Multiple logistic regression analysis indicated that residing in urban areas, parental education, monthly family income, and tooth brushing p<0.001) were identified as independent risk factors for oral cavity parasites. Conclusion: This study identified a notable prevalence of oral cavity parasites in children with ID in Lorestan province, Western Iran. It is imperative to recognize the primary risk factors associated with these parasites, particularly inadequate teeth brushing, in order to enhance public and oral health strategies for children with ID. Therefore, pediatric dental professionals should remain vigilant regarding these risk factors to effectively recognize and address oral health issues in this population, thereby mitigating the occurrence of oral diseases and infections.