Amebiasis Research Articles

Determination of liver enzyme levels in patients that infected with E.histolytica in Samarra city

The current study aimed to reveal the evaluation of the level of liver function in infected with the parasite Entamoeba histolytica, Blood samples were collected during the period from the beginning of August 2021 until the month of April 2022. The separated sera were kept at -20 degrees until use. Enzyme-linked immunosorbent adsorption (ELISA) technique was used to detect dysentery amoeba infection. ALP, ALT, AST concentrations were measured using Spectophotomertric method. In the current study, 250 samples were taken, and the number of samples infected with the dysentery amoeba parasite was 120 samples with a percentage of 48%, and the infection rate for sex was 85 samples infected with males by 59%, and about 35 samples were infected with 33% of females.The results of the current study showed a significant P≤0.05 increase in the levels of (GPT 4.464±54.214), , ALP 0.5542±6.7440 when compared with control group (7.14 ± 39.340 ) and 0.5699 ± 5.8061) There was an increase in the levels of (ALT 4.409±46.35), AST 7.19±62.44, ALP 0.42386±6.4563) when compared with control group (7.14±39.340, 9.75±54.16, 0.5699±5.8061 respectively.

EFFECT OF ENTAMOEBA HISTOLYTIC TO THE IMMUNE CHEMOKINES IN SAMARRA CITY

The current study aimed to reveal the evaluation of the level of some immune chemokines and liver function in infected with the parasite Entamoeba Histolytica. , Blood samples were collected during the period from the beginning of August 2021 until the month of April 2022. The separated sera were kept at -20 temperature until use. Enzyme-linked immunosorbent adsorption (ELISA) technique was used to detect dysentery amoeba infection. CCR2 and CCL2 levels were measured using ELISA technique, while ALP, ALT, AST and TSP were measured using Spectophotomertric method. In the current study, 250 samples were taken, and the number of samples infected with the dysentery amoeba parasite was 120 samples, at a rate of 48%, The results of the current study showed a significant increase of P≤0.05, as it was found that there were significant differences in the group with diarrhea only without the presence of the dysentery amoeba parasite, where there was an increase in the levels of ALT, AST, ALP and globulin compared to the control group. It was also found that there were no significant differences in AST levels between the group infected with the dysentery amoeba parasite compared to the control group.

Prevalence of Amoebic Dysentery Among Population of Wasit Province

A microscopic examination of multiple faeces samples of 600 persons was carried out in Al-Karama hospital,Wasit province from October 2009 to Feberuary 2010. Overall prevalence of Entamoeba histolytica was 60(10%). Prevalence increased rapidly in age group(1day-10year) and there were no real differences between males and females. Prevalence was higher among families who live in rural areas than those in urban areas. The highest infection rate was recorded in December and the lowest was recorded in October.

A tail-specific protease is required for Legionella pneumophila intracellular multiplication.

Legionella pneumophila is a Gram-negative bacterium found in natural and man-made water systems where it replicates within amoebas and ciliates. In humans, once inside the lungs, L. pneumophila replicates in alveolar macrophages and causes Legionnaires' disease, a severe pneumonia. The Icm/Dot type IVb secretion system is a major virulence factor required for intracellular multiplication. The Icm/Dot system allows the secretion of effectors into the cytoplasm of the host cell. These effectors modify host cell vesicular trafficking and prevent maturation of the phagosome. The innate immune response is crucial in restricting L. pneumophila proliferation. TNF-α is one of the major cytokines involved in this process as it renders macrophages more resistant to L. pneumophila infection and induces apoptosis of L. pneumophila-infected macrophages. Tail-specific proteases (Tsp) are involved in tolerating thermal stress and in virulence. We have previously characterized the Tsp encoded by L. pneumophila, showing that it is important for surviving thermal stress and for infection of amoeba when a temperature change occurs during infection. Here, we demonstrated that Tsp is required for intracellular multiplication in macrophages. Absence of tsp is associated with higher production of TNF-α by macrophages in response to L. pneumophila infection. This effect is independent of the Icm/Dot secretion system.

Evaluation of blood urea, serum creatinine and complete blood count for some Iraqi children with amoebiasis infection

Observation of urea, creatinine levels and complete blood count (CBC) with Iraqi children amoebiasis. This a cross-sectional prospective study was conducted among children with intestinal amoebiasis infection coming to the outpatient clinic at Al-Elwiya Paediatrics Teaching Hospital in Baghdad from the 1st of June 2018 until the end of November 2018. The study included 245 samples of children between the ages of 13 and 1 years, 83 Amoebian Enteritis children and 162 diarrhoeal children as a control group (diarrhoea and gastrointestinal disorders due to viral, bacterial, parasitic infections and any causative effects of diarrhoea or intestinal infections). The statistical analysis was carried out using the statistical package SPSS 23, to compare categorical and continuous data using the number, percentage, minimum, maximum, mean values with standard deviation, the Chi-square test and the T-test. Statistical significance has been achieved at P-value < 0.05. Patients with intestinal amoebiasis infected with diarrhoea and GIT disorder compared to control groups. Blood Urea, Serum creatinine and CBC showed a mean ± standard deviation, statistically significant differences at p-value < 0.05. In conclusion, Uraemia, high white blood cell count and a significant reduction in the haematocrit percentage due to the prolonged interval of disease infection with severe dehydration and used improper drugs.

Molecular diversity for Entamoeba histolytica virulence factor genes isolated from patients with amoebic dysentery

Entamoebahistolytica is a parasitic protozoan that caused amoebiasis, a diseases which infects the liver and colon and spread all over the world. E.histolytica is causing the death of human cells and leading to develop invasion, and ulceration of the intestine. However, amoebic liver abscess(ALA) considers as major common extraintestinal disease caused by E.histolytica.The aims of this study, was to detect E. histolytica by polymerase chain reaction (PCR), and to differentiate it from other amoeba species in a stool samples of human. Study the pathogenicity of E. histolytica in human through determining the presence of virulence factors like cysteine protease, amoebapore and Gal/lectin. A total of 100 stool samples were collected from patients with amoebic dysentery and then examined microscopically to confirm the presence of E. histolytica and other Entamoeba species, 72 ( 72%) out of 100 examined samples with E.histolytica were positive by PCR and 28(28%) were negative. The positive samples with PCR have been submitted to detect the virulence factors of E.histolytica by PCR and the results showed that the total of 72(100%)were positive with cysteine proteases gene, shown 62(86.11%) were positive with amoebapore and 65(90.27) were positive with lectin. Gene sequence for cysteine protease, amoebapore and lectin in local E. histolytica human isolates showed the nucleotide alignment similarity and substitution mutations in all of cysteine protease, amoebapore and Gal/GalNAc lectin. Phylogenetic tree analysis based on cysteine protease gene partial sequence in local E.histolytica human isolates that used for genetic variation analysis showed genetic closed related to NCBI-BLAST E.histolytica Japan strain (AK421547.1) at total genetic changes (0.0060-0.0010%), while Phylogenetic tree analysis based on amoebapore B gene partial sequence in local E.histolytica human isolates that used for genetic variation analysis showed genetic closed related to NCBI-BLAST E.histolytica, Germany strain (X76904.1) at total genetic changes (0.0060-0.0010%). Phylogenetic tree analysis based on Gal/GalNAc lectin gene partial sequence in local E. histolytica human isolates that used for genetic variation analysis showed genetic closed related to NCBI-BLAST E.histolytica Bangladesh strain (AF501280.1) at total genetic changes (0.0060-0.0020.

Evaluation of the Methods Used for the Detection of Entamoeba histolytica in Stool Samples of Patients with Diarrhea

Amoebic dysentery (amebiasis) is a parasitic infection caused by Entamoeba histolytica. The diagnosis of invasive amebiasis has traditionally been based on direct and stained microscopic examination of stool samples. Stool microscopy exhibits low sensitivity and it is difficult to distinguish E.histolytica cysts and trophozoites from cells such as leukocytes, macrophages and non-pathogenic Entamoeba species in the stool by microscopy. Therefore more sensitive and specific diagnostic methods such as enzyme linked immunosorbent assay (ELISA) tests which investigate the presence of E.histolytica-specific antigen in stool, and polymerase chain reaction (PCR) are being widely used. In this study it was aimed to study stool samples of the patients who applied with the clinical signs of amebiasis by using direct and permanent stained microscopy, E.histolytica adhesin antigen ELISA test and real-time PCR-based BD Max Enteric Parasite Panel (BD Max EPP) test and to evaluate the diagnostic values of these tests. A total of 546 faecal samples with blood and/or mucus were analyzed in the study. In these samples, the presence of E.histolytica was investigated by direct and permanent stained microscopy, E.histolytica adhesin antigen ELISA and BD Max EPP PCR. Of the samples 36.3% were suspected to contain E.histolytica/dispar/moshkovskii cyst and/or trophozoite by direct microscopic examination. Trichrome staining was performed on these samples and 49 samples were found suspicious for the presence of E.histolytica/dispar/moshkovskii cysts and/or trophozoites. The presence of E.histolytica and other Entamoeba species was not confirmed in 75.2% of the samples. BD Max EPP PCR and E.histolytica adhesin antigen ELISA tests were studied in 49 faecal samples that were suspected by trichrome staining. None of these samples were positive by ELISA. Forty-four samples were negative by PCR and invalid test results were obtained in five samples. In this study, E.histolytica was not detected in the patient population. The results of this study showed that microscopic examination alone is not sufficient for the detection of E.histolytica. It is concluded that it is necessary to use a more sensitive and specific also rapid diagnostic test such as E.histolytica-specific antigen detection test or PCR in the diagnosis of amebiasis to avoid misdiagnosis and unnecessary treatment of patients.

Formulation development and Evaluation of Colon Specific Delivery System of Anti- amoebic Drug (Tinidazole) using different Polysaccharide carriers

Amoebiasis is an infection of the large intestine caused by Entamoeba histolytica. Tinidazole is the preferred drug used in the treatment of intestinal amoebiasis. The conventional tablet dosage form provides minimal amount of drug in the colon with undesirable adverse effect due to variation in the transit time. Hence, to target the drug directly to the site of action in the colon, there is need to develop colon targeted drug delivery systems that will enhance the therapeutic drug level, increases the bioavailability of active medicament and reduce the dose of drug. The fast disintegrating core tablet of Tinidazole was prepared by direct compression method. The core tablet was evaluated for hardness, thickness, Friability, weight variation, drug content, in vitro disintegration time and in vitro drug release studies. Using the optimized formulation (C3), different formulations (F1-F6) of coated tablet was prepared by using different polymers like chitosan and xanthan gum. The formulated coated tablet were evaluated for various parameters like for hardness, thickness, Friability, weight variation, drug content, swelling study and in vitro dissolution study. The optimized formulation F3 shows 6.24% drug release in Stomach and Small Intestine and maximum drug release up to 96.53% in the colon. The kinetic data analysis of formulations indicated that it fits to Korsmeyer -Peppas Model and follows zero order release kinetics. The mechanism of drug release from the tablet followed non-Fickian (super case-11) transport. The stability studies of the optimized formulation were investigated as per ICH guidelines. There were no significant changes in thickness, hardness, weight variation, friability, drug content and in vitro drug release. These findings suggest that compression coated tablet using polysaccharide carriers provide more site specific delivery, reduced degradation and release of drug in stomach and small intestine.

Novel Antiamoebic Tyrocidine-Derived Peptide against Brain-Eating Amoebae.

Acanthamoeba castellanii (A. castellanii) can cause Acanthamoeba keratitis, a sight-threatening infection, as well as a fatal brain infection termed granulomatous amoebic encephalitis, mostly in immunocompromised individuals. In contrast, Naegleria fowleri (N. fowleri) causes a deadly infection involving the central nervous system, recognized as primary amoebic encephalitis, mainly in individuals partaking in recreational water activities or those with nasal exposure to contaminated water. Worryingly, mortality rates due to these infections are more than 90%, suggesting the need to find alternative therapies. In this study, antiamoebic activity of a peptide based on the structure of the antibiotic tyrocidine was evaluated against A. castellanii and N. fowleri. The tyrocidine-derived peptide displayed significant amoebicidal efficacy against A. castellanii and N. fowleri. At 250 μg/mL, the peptide drastically reduced amoebae viability up to 13% and 21% after 2 h of incubation against N. fowleri and A. castellanii., whereas, after 24 h of incubation, the peptide showed 86% and 94% amoebicidal activity against A. castellanii and N. fowleri. Furthermore, amoebae pretreated with 100 μg/mL peptide inhibited 35% and 53% A. castellanii and N. fowleri, while, at 250 μg/mL, 84% and 94% A. castellanii and N. fowleri failed to adhere to human cells. Amoeba-mediated cell cytopathogenicity assays revealed 31% and 42% inhibition at 100 μg/mL, while at 250 μg/mL 75% and 86% A. castellanii and N. fowleri were inhibited. Assays revealed inhibition of encystation in both A. castellanii (58% and 93%) and N. fowleri (73% and 97%) at concentrations of 100 and 250 μg/mL respectively. Importantly, tyrocidine-derived peptide depicted minimal cytotoxicity to human cells and, thus, may be a potential candidate in the rational development of a treatment regimen against free-living amoebae infections. Future studies are necessary to elucidate the in vivo effects of tyrocidine-derived peptide against these and other pathogenic amoebae of importance.

Identification of Multiple Domains of Entamoeba histolytica Intermediate Subunit Lectin-1 with Hemolytic and Cytotoxic Activities.

Galactose and N-acetyl-D-galactosamine-inhibitable lectin of Entamoeba histolytica have roles in the pathogenicity of intestinal amoebiasis. Igl1, the intermediate subunit lectin-1 of E. histolytica, has been shown to have both hemolytic and cytotoxic activities that reside in the C-terminus of the protein. To identify the amino acid regions responsible for these activities, recombinant proteins were prepared and used in hemolytic and cytotoxic assays. The results revealed that Igl1 has multiple domains with hemolytic and cytotoxic activities and that amino acids 787-846, 968-1028 and 1029-1088 are involved in these activities. The hemolytic activities of the fragments were partly inhibited by mannose, galactose and N-acetylgalactosamine, and glucose showed lower or negligible inhibitory effects for the activities. This is the first report of a protozoan protein with hemolytic and cytotoxic activities in multiple domains.

Molecular prevalence of Entamoeba species in a cohort of Egyptians

Background and objective: Intestinal and extraintestinal amebiasis is caused by the protozoan parasite Entamoeba (E.) histolytica, it is of considerable morbidity and mortality in developing countries. E.histoltica complex species includes E.histolytica, E. moshkovskii and E. dispar are morphologically indistinctable. The current study goal was to use molecular assays to detect the true prevalence of E. histolytica complex species among a cohort of Egyptians. Methods: A single stool specimen was collected from 133 patients, examined coproscopically before and after concentration. DNA was extracted from microscopically positive stool specimens for E. histolytica complex species were molecularly idenitified using multiplex PCR. Results: The coroscopic prevalence of intestinal parasites was 51.1% (68/133) of them 30 cases had E.histolytica complex (22.6%; 30/133). Among coproscopically positive samples, E. dispar was the most common parasite (63.3%; 19/30), followed by E. histolytica (23.4%; 7/30) and E. moshkovskii (13.3%; 4/30). There was statistical significance association between sociodemographic characteristics and Entamoeba species in asymptomatic individuals, while in the symptomatic individual, only age groups were stastically significant. Conclusion: E.dispar is the predominant Entamoeba species among studied individuals. There is a need for molecular diagnosis of Entamoeba to determine the true prevalence of E. histolytica and avoid overmedication.

Assessment of Entamoeba histolytica Coproantigen and Specific Salivary IgA in Relation to Real-Time PCR for Detection of Intestinal Amoebiasis

Assessment of Entamoeba histolytica Coproantigen and Specific Salivary IgA in Relation to Real-Time PCR for Detection of Intestinal Amoebiasis

Diagnostic and molecular study of Entamoeba dispar in Nineveh Province, Iraq

Amoebiasis is one of the important parasitic intestinal diseases that is widespread in mosul and all Iraq, responsible for many deaths, especially in children, caused by an elementary parasite Entamoeba histolytica, and there are other species of the genus Amoeba, which are not pathological, such as Entamoeba dispar., and because of the similarity between the two species in morphology, it has become necessary to use the accurate diagnosis of amoebiasis, this study was conducted for the period between October 2021 and until April 2022, included the examination of 96 stool samples from the patients in Nineveh Governorate hospitals and some private medical laboratories whose ages ranged from less than one year to 50 years and for both genders, during this study, microscopic and molecular diagnostic methods were used for two species of the genus Amoeba, as the microscopic examination was carried out using a direct wet smear to detect the stages of the trophozoite parasite, the cyst, or both. The study showed that there was an infection rate of 33.3%, with a rate of (32) of the stool samples diagnosed microscopically out of a total of 96 samples among the examined persons, while the results of using molecular diagnosis using polymerase chain reaction technology showed for 50 stool samples out of 96 samples (32 positive samples and 18 negative samples), the presence of amoeba infection was observed by 38%. In addition, two species of amoeba were diagnosed in the examined samples, the highest rate of infection was for Entamoeba histolytica, followed by Entamoeba dispar, at a rate of 52.63% and 47.36%, respectively. In addition, the Gene sequencer for the species E.dispar was performed. Which was isolated for the first time in Nineveh Governorate and registered in the National Center for Genetic Information with serial number 692812.

Common Illness with an Uncommon Course- In Adolescents use HEADSS

Adolescence is a period of emotional instability, during which high risk behavior including experimentation with drugs is common. Clinical manifestations of substance abuse and its withdrawal may mimic common illnesses. We report an adolescent who presented with acute dysentery with unusual course. HEADSS screening revealed psychosocial stressors leading to cannabis addiction. The child was diagnosed to have cannabis withdrawal. He was referred to the de-addiction clinic. HEADSS is a useful tool to identify risk factors and protective factors in adolescents. HEADSS screening during each interaction and early intervention when needed must be an essential component of adolescent health care.

An Unusual U2AF2 Inhibits Splicing and Attenuates the Virulence of the Human Protozoan Parasite Entamoeba histolytica.

E. histolytica is the etiological agent of intestinal amebiasis and liver abscesses, which still poses public health threat globally. Metronidazole is the drug of choice against amebiasis. However, metronidazole-resistant amoebic clinical isolates and strains have been reported recently, challenging the efforts for amebiasis eradication. In search of alternative treatments, E. histolytica transcriptomes have shown the association of genes involved in RNA metabolism with the virulence of the parasite. Among the upregulated genes in amoebic liver abscesses are the splicing factors EhU2AF2 and a paralog of EhSF3B1. For this reason and because EhU2AF2 contains unusual KH-QUA2 (84KQ) motifs in its lengthened C-terminus domain, here we investigated how the role of EhU2AF2 in pre-mRNA processing impacts the virulence of the parasite. We found that 84KQ is involved in splicing inhibition/intron retention of several virulence and non-virulence-related genes. The 84KQ domain interacts with the same domain of the constitutive splicing factor SF1 (SF1KQ), both in solution and when SF1KQ is bound to branchpoint signal RNA probes. The 84KQ–SF1KQ interaction prevents splicing complex E to A transition, thus inhibiting splicing. Surprisingly, the deletion of the 84KQ domain in EhU2AF2 amoeba transformants increased splicing and enhanced the in vitro and in vivo virulence phenotypes. We conclude that the interaction of the 84KQ and SF1KQ domains, probably involving additional factors, tunes down Entamoeba virulence by favoring intron retention.

The Role of Acetate Kinase in the Human Parasite Entamoeba histolytica.

The human parasite Entamoeba histolytica, which causes approximately 100 million cases of amoebic dysentery each year, relies on glycolysis as the major source of ATP production from glucose as it lacks a citric acid cycle and oxidative phosphorylation. Ethanol and acetate, the two major glycolytic end products for E. histolytica, are produced at a ratio of 2:1 under anaerobic conditions, creating an imbalance between NADH production and utilization. In this study we investigated the role of acetate kinase (ACK) in acetate production during glycolysis in E. histolytica metabolism. Analysis of intracellular and extracellular metabolites demonstrated that acetate levels were unaffected in an ACK RNAi cell line, but acetyl-CoA levels and the NAD+/NADH ratio were significantly elevated. Moreover, we demonstrated that glyceraldehyde 3-phosphate dehydrogenase catalyzes the ACK-dependent conversion of acetaldehyde to acetyl phosphate in E. histolytica. We propose that ACK is not a major contributor to acetate production, but instead provides a mechanism for maintaining the NAD+/NADH balance during ethanol production in the extended glycolytic pathway.

Factors Associated with Amoebic Dysentery in Captive Non-Human Primates of The Mefou Primate Sanctuary

Primates are known to harbour different gastrointestinal parasite species that affect their survival and reproductive activity. Entamoeba histolytica infects humans predominantly as well as non-human (NH) primates causing amoebiasis. Amoebic dysentery is common amongst non-human primates in captive sites. The purpose of the study was to evaluate the risk factors associated with the prevalence of amoebic dysentery in captive NH primates of the Mefou Primate Sanctuary (MPS) in Cameroon. The faecal samples of 277 NH primates were analysed by qualitative (direct smear and evergreen kit sedimentation technique) coprology from January to July 2019. Factors such as habitat type, keeper’s age and level of education were assessed to evaluate their effect on the prevalence of amoebic dysentery. The prevalence of Entamoeba histolytica was 46.2% (P=0,079 and χ2=18.13). E histolytica was the most predominant parasite species detected and it was present in all instances of mixed parasite infections. Amongst the NH primates positive for E. histolytica, 28 suffered from amoebic dysentery giving a prevalence of 21.9% and one of the infected primates (Cercopithecus pogonias) actually died from amoebic dysentery. The prevalence of mixed infections was significantly lower amongst NH primates living in outdoor enclosures compared to those living in other habitat types (P=0.015, χ2=10.46). The rate of re-infection was significantly higher amongst NH primates under the care of keepers with the lowest (primary school) level of education (P=0.001, χ2=13.09) on one hand and of older keepers (≥50s) (P = 0.008, χ2=13.708) on the other hand. This study shows that amoebic dysentery is a cause for a major concern in the MPS. The housing conditions of the NH primates at the sanctuary, the keeper’s age and level of education were important factors that influenced the prevalence of amoebic dysentery. Adequate measures should be put in place to address the situation.

A mini-review of pharmacological and economical values of Funtumia africana

Funtumia africana belongs to the family Apocynaceae. It is a fast growing tree found in green forests and forest reserves in West Africa. The plant has a long history of use in traditional medicine to treat and manage various diseases such as diabetes, cancer, burns, amoebic dysentery, urinary incontinence, bacterial infections and malaria. It is used economically for the production of furniture, match sticks and firewood. This review paper presents the botanical description, pharmacological uses and economic importance of F. africana.

A rare case of extraintestinal amebiasis

BackgroundAmoebiasis is caused by the protozoan Entamoeba histolytica, which is a rare infectious disease in developed countries. If the trophozoites enter the blood, it can spread through the body, such as brain, and lungs. Cases of simultaneous infection of multiple organs are extremely rare.Case presentationHere we report a case of simultaneous infection of amoeba in pulmonary pleura, urinary system and central nervous system. Although the patient received anti amoeba treatment, the prognosis of the patient was poor.ConclusionsIn this patient, multiple extraintestinal amebic infections in the absence of clinically confirmed intestinal amebiasis or amebic liver abscess are rare and pose diagnostic challenges. The disseminated amebiasis has significantly increased the mortality. Early diagnosis and appropriate treatment may reduce the mortality of disseminated amebiasis.

Polyaniline (PANI)-conjugated tungsten disulphide (WS2) nanoparticles as potential therapeutics against brain-eating amoebae.

Brain-eating amoebae, including Acanthamoeba castellanii and Naegleria fowleri, are the causative agents of devastating central nervous system infections with extreme mortality rates. There is an indisputable urgency for the development of effective chemotherapeutic agents for the control of these diseases that are increasing in incidence. Here, we evaluated the anti-amoebic potential of polyaniline:tungsten disulphide (PANI:WS2) nanocomposite against the infective trophozoite and cyst stages of N. fowleri and A. castellanii. Throughout these evaluations, significant viability inhibition was noted when 100µg/mL of PANI:WS2 was employed at its 1:5 formulation. These effects were studied to be due to increased levels of reactive oxygen species (ROS) as visualised through fluorescence microscopy. Furthermore, field emission scanning electron microscopy (FE-SEM) analysis pictured disruption to amoeba morphology. The host-cell cytotoxicity of the nanocomposite (PANI:WS2) was studied to be negligible, making it an attractive avenue in the pursuit for effective treatments for brain-eating amoeba infections. KEY POINTS: • Synthesis of polyaniline:tungsten disulphide (PANI:WS2) nanocomposite. • Anti-amoebic potential of PANI:WS2 nanocomposite. • PANI:WS2 nanocomposites are promising anti-amoebic agents in vitro.