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Objective: This study aimed to retrospectively evaluate the prevalence of protozoan parasites in stool samples collected from patients presenting with various gastrointestinal complaints to the Medical Parasitology Laboratory of Kafkas University Research and Application Hospital between 2019 and 2022., Methods: Stool samples were initially examined using the native-Lugol method for the detection of protozoan parasites, followed by the formol-ethyl acetate sedimentation method, Giemsa, and carbol fuchsin staining methods. Specific immunochromatographic card tests were used for the diagnosis of Entamoeba histolytica, Cryptosporidium spp., and Giardia intestinalis ., Results: Of the 2.267 stool samples examined over the four-year period from January 2019 to December 2022, 7.63% were found to contain one or more protozoan parasites. Among these parasites, Entamoeba histolytica was detected at the highest rate of 4.06%. The other parasite species were identified as follows: Blastocystis spp. 1.15%, Entamoeba spp. and Entamoeba coli each 0.52%, Giardia intestinalis 0.48%, Endolimax nana 0.17%, and Entamoeba histolytica/dispar 0.08%., Conclusion: This study indicates that despite a decrease in the prevalence of intestinal protozoan infections in the Kars region, these infections remain a significant public health issue. Therefore, improvements in hygiene and sanitation conditions, increased public health education, and the widespread implementation of early diagnosis and treatment methods are necessary. Special measures should be taken to protect vulnerable groups, particularly children and the elderly., Competing Interests: Conflict of Interest: No conflict of interest was declared by the authors.

The development of rapid, accurate, and efficient detection methods for protozoan parasites can substantially control the outbreak of protozoan parasites infection, which poses a threat to global public health. Idealistically, electrochemical biosensors would be able to overcome the limitations of current detection methods due to their simplified detection procedure, on-site quantitative analysis, rapid detection time, high sensitivity, and portability. The objective of this scoping review is to evaluate the current state of electrochemical biosensors for detecting protozoan parasites. This review followed the most recent Preferred Reporting Items for Systematic Review and Meta-Analyses extension for Scoping Reviews (PRISMA-ScR) recommendations. Using electrochemical biosensor and protozoan parasite keywords, a literature search was conducted in PubMed, Scopus, Web of Science, and ScienceDirect on journals published between January 2014 and January 2022. Of the 52 studies, 19 were evaluated for eligibility, and 11 met the review's inclusion criteria to evaluate the effectiveness and limitations of the developed electrochemical biosensor platforms for detecting protozoan parasite including information about the samples, biomarkers, bioreceptors, detection system platform, nanomaterials used in fabrication, and limit of detection (LoD). Most electrochemical biosensors were fabricated using conventional electrodes rather than screen-printed electrodes (SPE). The range of the linear calibration curves for the developed electrochemical biosensors was between 200 ng/ml and 0.77 pM. The encouraging detection performance of the electrochemical biosensors demonstrate their potential as a superior alternative to existing detection techniques. On the other hand, more study is needed to determine the sensitivity and specificity of the electrochemical sensing platform for protozoan parasite detection.

Stool examination using microscopy was the traditional method for the diagnosis of intestinal parasites. Recently, the use of molecular tests to identify stool protozoa has become the main tool used in most clinical laboratories in Israel. This study aimed to evaluate the prevalence of intestinal parasites in Israel and to compare this prevalence in laboratories that use molecular tests vs a laboratory that uses microscopy. Samples collected from January to October 2021 at seven laboratories were analyzed by real-time PCR (RT-PCR) or by microscopy. The multiplex panel included the following pathogens: Giardia lamblia , Entamoeba histolytica , Cryptosporidium spp. , Cyclospora , Dientamoeba fragilis , and Blastocystis spp . Overall, 138,415 stool samples were tested by RT-PCR and 6,444 by microscopy. At least one protozoa species was identified in 28.4% of the PCR-tested samples compared to 4.6% of the microscopy-tested samples. D. fragilis was the most common PCR-identified species (29%). D. fragilis , G. lamblia , and Cryptosporidium spp . were mainly found in pediatric population, while Blastocystis spp . was most prevalent among adults ( P < 0.001). In a sub-cohort of 21,480 samples, co-infection was found in 4,113 (19.15%) samples, with Blastocystis spp . and D. fragilis being the most common (14.9%) pair. Molecular stool testing proved more sensitive compared to microscopy. D. fragilis was the most commonly detected pathogen. The above profile was identified during the COVID pandemic when traveling was highly restricted and most likely represents the locally circulating protozoa., Importance: This study sheds light on the prevalence of stool parasites in Israel. Additionally, this study indicates that the shift from microscope analysis to molecular tests improved protozoa diagnosis., Competing Interests: The authors declare no conflict of interest.

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., Competing Interests: Declaration of conflicting interestsThe authors declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.

In this case report, we address the diagnostic challenges and clinical implications of severe infection with Lophomonas blattarum in a patient initially suspected of experiencing long COVID symptoms. We describe the patient's medical history, initial symptoms, diagnostic tests, and treatment. A female patient with diabetes in her early 60s presented with severe shortness of breath and was initially diagnosed with diabetic ketoacidosis (DKA). After resolution of her DKA symptoms, persistent respiratory issues led to a COVID-19 test, which was negative. A chest computed tomography scan revealed abnormalities, prompting bronchoscopy and bronchoalveolar lavage fluid analysis, which confirmed the presence of L. blattarum . Notably, the protozoan remained mobile and viable even after a 4-day transport at ambient temperature. This case emphasizes the importance of considering alternative diagnoses and improving awareness about L. blattarum infection in patients with respiratory symptoms, for timely and accurate management., Competing Interests: Declaration of conflicting interestThe authors declare that there is no conflict of interest.

Human infections with the protozoan Lophomonas have been increasingly reported in the medical literature over the past three decades. Initial reports were based on microscopic identification of the purported pathogen in respiratory specimens. Later, a polymerase chain reaction (PCR) was developed to detect Lophomonas blattarum , following which there has been a significant increase in reports. In this minireview, we thoroughly examine the published reports of Lophomonas infection to evaluate its potential role as a human pathogen. We examined the published images and videos of purported Lophomonas, compared its morphology and motility characteristics with host bronchial ciliated epithelial cells and true L. blattarum derived from cockroaches, analyzed the published PCR that is being used for its diagnosis, and reviewed the clinical data of patients reported in the English and Chinese literature. From our analysis, we conclude that the images and videos from human specimens do not represent true Lophomonas and are predominantly misidentified ciliated epithelial cells. Additionally, we note that there is insufficient clinical evidence to attribute the cases to Lophomonas infection, as the clinical manifestations are non-specific, possibly caused by other infections and comorbidities, and there is no associated tissue pathology attributable to Lophomonas . Finally, our analysis reveals that the published PCR is not specific to Lophomonas and can amplify DNA from commensal trichomonads. Based on this thorough review, we emphasize the need for rigorous scientific scrutiny before a microorganism is acknowledged as a novel human pathogen and discuss the potential harms of misdiagnoses for patient care and scientific literature., Competing Interests: The authors declare no conflict of interest.

Globally, over 3.5 billion people are infected with intestinal parasites each year, resulting in over 200,000 deaths. Three of the most common protozoan pathogens that affect the gastrointestinal tract of humans are Cryptosporidium spp., Giardia intestinalis , and Entamoeba histolytica . Other protozoan agents that have been implicated in gastroenteritis in humans include Cyclospora cayetanensis , Dientamoeba fragilis , Blastocystis hominis , and the microsporidia Enterocytozoon bieneusi and Encephalitozoon intestinalis . Genetic Signatures previously developed a 3base™ multiplexed Real-Time PCR (mRT-PCR) enteric protozoan kit (EP001) for the detection of Giardia intestinalis / lamblia / duodenalis , Cryptosporidium spp., E. histolytica , D . fragilis , and B. hominis . We now describe improvements to this kit to produce a more comprehensive assay, including C. cayetanensis , E. bieneusi , and E. intestinalis , termed EP005. The clinical performance of EP005 was assessed using a set of 380 clinical samples against a commercially available PCR test and other in-house nucleic acid amplification tests where commercial tests were not available. All methods provided at least 90% agreement. EP005 had no cross-reactivity against 82 organisms commonly found in the gut. The EP005 method streamlines the detection of gastrointestinal parasites and addresses the many challenges of traditional microscopic detection, resulting in cost savings and significant improvements in patient care.

Annexins (ANXs) exert different functions in cell biological and pathological processes and are thus known as double or multi-faceted proteins. These sophisticated proteins might express on both parasite structure and secretion and in parasite-infected host cells. In addition to the characterization of these pivotal proteins, describing their mechanism of action can be also fruitful in recognizing their roles in the pathogenesis of parasitic infections. Accordingly, this study presents the most prominent ANXs thus far identified and their relevant functions in parasites and infected host cells during pathogenesis, especially in the most important intracellular protozoan parasitic infections including leishmaniasis, toxoplasmosis, malaria and trypanosomiasis. The data provided in this study demonstrate that the helminth parasites most probably express and secret ANXs to develop pathogenesis while the modulation of the host-ANXs could be employed as a crucial strategy by intracellular protozoan parasites. Moreover, such data highlight that the use of analogs of both parasite and host ANX peptides (which mimic or regulate ANXs physiological functions through various strategies) might suggest novel therapeutic insights into the treatment of parasitic infections. Furthermore, due to the prominent immunoregulatory activities of ANXs during most parasitic infections and the expression levels of these proteins in some parasitic infected tissues, such multifunctional proteins might be also potentially relevant as vaccine and diagnostic biomarkers. We also suggest some prospects and insights that could be useful and applicable to form the basis of future experimental studies., Competing Interests: Declaration of Competing Interest The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023. Published by Elsevier B.V.)

Lophomonas blattarum is an anaerobic protozoan living in the intestine of cockroaches and house dust mites, with ultramicroscopic characteristics such as the presence of a parabasal body, axial filament, and absence of mitochondria. More than 200 cases of Lophomonas infection of the respiratory tract have been reported worldwide. However, the current diagnosis of such infection depends only on light microscopic morphological findings from respiratory secretions. In this study, we attempted to provide more robust evidence of protozoal infection in an immunocompromised patient with atypical pneumonia, positive for Lophomonas-like protozoal cell forms. A direct search of bronchoalveolar lavage fluid via polymerase chain reaction (PCR), transmission electron microscopy (TEM), and metagenomic next-generation sequencing did not prove the presence of protozoal infection. PCR results were not validated with sufficient rigor, while de novo assembly and taxonomic classification results did not confirm the presence of an unidentified pathogen. The TEM results implied that such protozoal forms in light microscopy are actually non-detached ciliated epithelial cells. After ruling out infectious causes, the patient's final diagnosis was drug-induced pneumonitis. These findings underscore the lack of validation in the previously utilized diagnostic methods, and more evidence in the presence of L. blattarum is required to further prove its pathogenicity.

Background: Cancer is a category of diseases that cause an individual's immune system to become suppressed. In a case-control study, the current study aims to detect the frequency of intestinal parasites and related risk factors in children with cancer., Methods: Stool samples were collected from 178 children with cancers (cases) and 150 cancer-free children (controls) who sought treatment for diarrheal episodes at nearby hospitals. Samples were processed by direct smear examination, concentration technique, permanent staining by Lugol's iodine, modified Ziehl-Neelsen, modified trichrome, and chromotrope 2R stains., Results: The overall prevalence of intestinal parasites was 7.3% (24/328), with non-statistically significant differences between cases (7.8%; 14/178) and controls (6.6%; 10/150). Children with leukemia had a higher infection rate (9%; 9/100) than children with lymphoma (6.9%; 3/43) or solid tumors (5.7%; 2/35). Blastocystis sp. (3.3%) was the most common intestinal parasite found in cases, followed by Cryptosporidium sp. (2.2%), Giardia lamblia (1.6%), and Microsporidia sp. (0.5%). For all parasites, no statistical difference was found between the two groups. (p > 0.05). Male gender, young age, non-bottled water use, travel to parasite-endemic areas, living in an urban area, and infrequent hand washing were all associated with intestinal parasitosis, with non-statistical significance observed between the two groups. In children with cancer, intestinal parasites were found to be significantly associated with chronic (p = 0.04) and severe (p = 0.03) diarrhea., Conclusions: Children with cancer, particularly those with hematological cancers, should be screened for intestinal parasites on a regular basis and treated for their overall health.

Background: Enteric parasite infections are underestimated due to the limited sensitivity and specificity of microscopy, which remains the diagnostic gold standard in routine clinical practice. This could be a major problem in high-income countries, where the burden of parasitic diseases is low. In recent years, Multiplex Real-Time polymerase chain reaction (RT-PCR) based methods have been implemented. Therefore, the aim of this study was to evaluate the prevalence of four enteric protozoan species detected by RT-PCR in non-native children in Italy, and to describe their clinical characteristics., Methods: Adopted and immigrant children, evaluated for migration health assessment between 2017 and 2020 in a tertiary care children's hospital in Italy, were enrolled. Molecular analysis for Giardia lamblia, Dientamoeba fragilis, Blastocystis hominis, and Entamoeba histolytica, was conducted by in-house RT-PCR., Results: Overall, 209 children were enrolled and 70% of them resulted positive by RT-PCR for at least one enteric parasite. B. hominis (47.8%) was the most commonly identified protozoa, followed by D. fragilis (44.5%). Co-infections with multiple pathogens were detected in 35.4% of the samples. Almost 80% of parasite-positive children were asymptomatic and the most common symptom was flatulence (60.7% of symptomatic children). Eosinophils were significantly increased in RT-PCR positive children compared to the negative ones and children with D. fragilis presented the highest eosinophils count., Conclusions: The In-house Multiplex RT-PCR assay provides a valid molecular detection system for selected enteric parasites. This novel and accurate diagnostic method can help in increasing the detection rate of parasite infection, especially in high-risk population., (Copyright © 2021. Published by Elsevier Ltd.)

Neospora caninum, Toxoplasma gondii and Hammondia spp. are coccidian parasites similar in morphology. Molecular techniques are necessary to detect parasite DNA isolated from stool samples in wild canids because they were reported as definitive hosts of N. caninum life cycle. The objective of this study was to develop a highly sensitive and accurate molecular method for the identification of coccidian Apicomplexa parasites in crab-eating fox (Cerdocyon thous) and pampas fox (Lycalopex gymnocercus). Tissue samples from road-killed animals (pampas fox = 46, crab-eating fox = 55) and feces (pampas fox = 84, crab-eating fox = 2) were collected, and species were diagnosed through molecular assay. PCR was used for the amplification of a fragment of the coccidian Apicomplexa nss-rRNA gene. Additionally, we developed a novel real-time PCR TaqMan™ probe approach to detect T. gondii- Hammondia spp. and N. caninum. This is the first report of N. caninum DNA in pampas fox feces (n = 1), thus it was also detected from pampas fox tissues (n = 1). Meanwhile, T. gondii was found in tissues of pampas (n = 1) and crab-eating (n = 1) foxes and H. triffittae in one crab-eating fox tissue. Despite the low percentage (2.5%) of positive samples, the molecular method developed in this study proved to be highly sensitive and accurate allowing to conduct an extensive monitoring analysis for these parasites in wildlife., (© 2021. The Author(s), under exclusive licence to Springer Nature B.V.)

Background: Metagenomic next generation sequencing (mNGS) is becoming increasingly available for pathogen detection directly from clinical specimens. These tests use target-independent, shotgun sequencing to detect potentially unlimited organisms. The promise of this methodology to aid infection diagnosis is demonstrated through early case reports and clinical studies. However, the optimal role of mNGS in clinical microbiology remains uncertain., Content: We reviewed studies reporting clinical use of mNGS for pathogen detection from various specimen types, including cerebrospinal fluid, plasma, lower respiratory specimens, and others. Published clinical study data were critically evaluated and summarized to identify promising clinical indications for mNGS-based testing, to assess the clinical impact of mNGS for each indication, and to recognize test limitations. Based on these clinical studies, early testing recommendations are made to guide clinical utilization of mNGS for pathogen detection. Finally, current barriers to routine clinical laboratory implementation of mNGS tests are highlighted., Summary: The promise of direct-from-specimen mNGS to enable challenging infection diagnoses has been demonstrated through early clinical studies of patients with meningitis or encephalitis, invasive fungal infections, community acquired pneumonia, and other clinical indications. However, the proportion of patient cases with positive clinical impact due to mNGS testing is low in published studies and the cost of testing is high, emphasizing the importance of improving our understanding of 'when to test' and for which patients mNGS testing is appropriate., (© American Association for Clinical Chemistry 2020. All rights reserved. For permissions, please email: journals.permissions@oup.com.)

Globalization has contributed to the emergence of specific parasitic diseases in novel geographical areas, and in these regions, these infections in travelers and immigrants may cause a considerable burden of disease. Timely diagnosis and treatment of protozoan infections to decrease mortality and prevent associated complications are essential. In this respect, the increased availability of specific DNA-detection procedures has improved the diagnosis of many imported parasitic infections. Travelers and immigrants with associated comorbidities or immunosuppression may pose a special challenge regarding management. An updated review of the main protozoan infections in mobile populations (malaria, Chagas disease, leishmaniasis, enteric protozoan infections) is provided, focusing on the changing epidemiology of these diseases, recent developments in diagnosis and management and the possibility of local transmission of imported infections.

Background: Nucleic acid-based methods are increasingly used for screening of gastrointestinal parasites. Microscopy is still used and Swedish routine protocol consists of formalin ethyl-acetate concentration and do not include screening for trophozoites or Cryptosporidium spp. This study aimed to compare detection with the Swedish routine microscopy method to an extended method that includes screening for trophozoites and Cryptosporidium. Furthermore, we also developed a method for DNA recovery from SAF-fixed faecal samples and compared the real-time PCR detection of Giardia intestinalis, Dientamoeba fragilis , Cryptosporidium spp., Entamoeba histolytica and Entamoeba dispar from SAF-fixed and unpreserved faecal samples. PCR results were then compared with microscopy results. Methods: SAF-fixed and unpreserved faecal samples from 1000 patients at the Clinical microbiology laboratory in Region Jönköping County, Sweden, were included. Samples were analysed with routine formalin ethyl-acetate concentration, wet mounts from both concentrated and unconcentrated samples, Ziehl-Neelsen staining on patients with certain symptoms and real-time PCR. Results: We found a significant higher detection rate of parasites with the extended microscopy method compared to the Swedish routine microscopy method when SAF-fixed samples were used. The detection rate with real-time PCR in SAF-fixed samples was equal to the detection rate in unpreserved samples. There was no significant difference in detection comparing extended microscopy and real-time PCR. Conclusion: In conclusion, this study showed that the extended microscopy method increased detection of intestinal protozoa with detection of both trophozoites and Cryptosporidium spp. We also showed that SAF-fixative can be used for detection of parasite-DNA with real-time PCR.

Infections causes by parasites of the gastrointestinal tract are a global public health problem. In industrialised countries, their particular epidemiological (low general prevalence of enteroparasites), economic (high labour costs) and clinical characteristics (constant increase in the number of samples and diagnostic determinations to be performed) have led molecular techniques to progressively replace conventional microscopy as the first-line diagnostic method of these pathogens in modern clinical laboratories. PCR-based techniques, particularly those developed for the simultaneous detection of the various agents that can cause the same infectious disease (syndromic diagnosis), already represent a cost-effective option that allow process automisation, workflow optimisation, and comparison of results among different laboratories, and facilitate accreditation of diagnostic procedures. This review clearly and concisely discusses the current situation of the molecular diagnosis of the main species of intestinal parasites in humans, particularly the enteric protozoans causing diarrhoea (Cryptosporidium spp., Giardia duodenalis, Entamoeba histolytica), the most important members the Microsporidia phyla (Enterocytozoon bieneusi) and Stramenopiles phyla (Blastocystis sp.), as well as the helminths transmitted by soil (Ancylostoma spp., Ascaris lumbricoides, Necator americanus, Strongyloides stercoralis and Trichuris trichiura) and food (Anisakis spp., Clonorchis sinensis, Fasciola spp., Taenia solium, and Trichinella spiralis). Special attention is paid to the description of available techniques and formats, to their diagnostic benefits and the most widely used genetic markers for their detection, both in clinical laboratories and genotyping in referral and research centres., (Copyright © 2020. Publicado por Elsevier España, S.L.U.)

Purpose of Review: This article reviews common infectious optic neuropathies, focusing on the more common and globally important entities., Recent Findings: Novel infections continue to emerge and drift geographically over time; not infrequently, these have important neurologic or ocular features. Malarial retinal findings comprise a relatively specific set of findings and serve as an invaluable aid in the diagnosis of cerebral malaria. Therapy continues to evolve and is best formulated in concert with an infectious disease expert., Summary: Infectious optic neuropathies are less common than inflammatory or ischemic optic neuropathies; may present with varied, overlapping, and nonspecific clinical appearances; and comprise an important differential consideration demanding specific therapy.

These updated guidelines from the Infectious Diseases Community of Practice of the American Society of Transplantation review the diagnosis, prevention, and management of tissue and blood protozoal infections in the pre- and post-transplant period. Significant new developments in the field have made it necessary to divide the previous single guideline published in 2013 into two sections, with the intestinal parasites separated from this guideline devoted to tissue and blood protozoa. The current update reflects the increased focus on donor screening and risk-based recipient monitoring for parasitic infections. Increased donor testing has led to new recommendations for recipient management of Toxoplasma gondii and Trypanosoma cruzi. Molecular diagnostics have impacted the field, with access to rapid diagnostic testing for malaria and polymerase chain reaction testing for Leishmania. Changes in Babesia treatment regimens in the immunocompromised host are outlined. The risk of donor transmission of free-living amebae infection is reviewed. Changing immigration patterns and the expansion of transplant medicine in developing countries has contributed to the recognition of parasitic infections as an important threat to transplant outcomes. Medications such as benznidazole and miltefosine are now available to US prescribers as access to treatment of tissue and blood protozoa is increasingly prioritized., (© 2019 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

Human stool contains a myriad of microorganisms, of which the vast majority are nonpathogenic and represent an important component of the healthy microbiome. The increasing use of molecular techniques has allowed the rapid identification of bacteria, viruses and parasites in human stool. This review focuses on the 3 main classes of parasite responsible for human disease, helminths, protozoa and ectoparasites, and highlights the importance of differentiating between pathogenic and nonpathogenic parasites.

Objectives: A stool sample is the sample of choice for microbiological testing of enteric pathogens causing diarrhoea, but a rectal swab can be a more practical alternative. A prospective observational study was performed to evaluate the diagnostic performance of flocked rectal swab specimens using the syndromic molecular approach to determine the aetiology of diarrhoea in adults., Methods: We compared the performance of rectal swabs with stool samples as the reference standard in determining viral, bacterial and protozoal pathogens using real-time multiplex PCR as well as standard stool culture. Paired samples of stool and rectal swab specimens were collected from 304 adult patients with diarrhoea, presented at the Department of Infectious Diseases, University Medical Centre Ljubljana, between June 2016 and August 2017., Results: Overall sensitivity of rectal swab samples in the syndromic molecular approach was 83.2% (95% CI 77.2%-88.1%). Pathogen group-specific analysis of rectal swabs showed sensitivity of 65.6% (95% CI 52.7%-77.1%) for viruses and 57.1% (95% CI 28.9%-82.3%) for parasites. For bacteria, sensitivity was 86.5% (95% CI 79.5%-91.8%) when PCR was performed and 61.4% (95% CI 52.4%-69.9%) when culture for bacteria was performed. Mean threshold cycle (Ct) values for most pathogens were higher in rectal swab specimens than in stool specimens., Conclusions: Our results indicate that rectal swabs can be used in the diagnosis of diarrhoea in adults when stool specimens are not available or when rapid aetiological determination is needed. However, rectal swabs should be analysed using a molecular approach. The mean Ct value for most pathogens is higher in rectal swab specimens than in stool specimens., (Copyright © 2018 European Society of Clinical Microbiology and Infectious Diseases. Published by Elsevier Ltd. All rights reserved.)

Paramyxean parasites in the genus Marteilia deteriorate digestive tissues of the host organisms, resulting in mortality of oysters, cockles, and mussels. Most reports of infection by Marteilia spp. are from Europe, while a new species of Marteilia was identified recently in Japan. Here, we report a previously unidentified species in the genus Marteilia from digestive diverticula of Manila clam Ruditapes philippinarum from the south coast of Korea. Prevalence of the parasite was low, 0.5-3.3% in the study sites. We characterized this species using light and transmission electron microscopy (TEM), and analyzed the 18S rDNA sequence. Light microscopy revealed the sporulation process from uninucleated stage to spore in the epithelial tissues of the digestive gland. TEM revealed that the parasites produced four secondary cells containing four tri-cellular spores. An electron-dense haplosporosome-like structure and striated inclusions were evident in the spore and the primary cells, respectively, while refringent granules were rarely observed in the secondary cells. Phylogenetic analyses of the 18S rDNA sequence placed this isolate in the genus Marteilia, although it is not identical to other known species in the genus. Based on morphological and molecular characters, we describe this species as Marteilia tapetis sp. nov., the second Marteilia species reported parasitizing Manila clams in Asian waters., (Copyright © 2019. Published by Elsevier Inc.)

The aim of this study was to evaluate the association between a longer sedimentation time, the reading of a larger number of slides, and the collection of multiple samples on the efficiency of the spontaneous sedimentation technique. Twenty-two patients with a previous parasitological exam positive for intestinal protozoa were recruited to collect new fecal samples (3 samples per patient) before the beginning of antiparasitic treatment. All collected fecal samples were used for spontaneous sedimentation and centrifuge-flotation techniques. Of these, all 22 patients were positive based on spontaneous sedimentation, and 59.1% (13/22) based on centrifuge flotation. The number of samples and the number of slides analyzed by spontaneous sedimentation influenced the number of positive cases. The modifications applied to the spontaneous sedimentation technique increased its performance in protozoa diagnosis.

The frequency of bronchopulmonary protozoan infections has raised due to increased number of immunosuppressed patients in recent years. One of them is Lophomonas blattarum which is a multi-flagellated protozoan parasite of termites and several cockroach species. The drug regimens commonly used in bronchopulmonary infections are not effective against L. blattarum . Therefore, rapid and accurate diagnosis of L. blattarum infection is of great importance in the treatment success. The laboratory diagnosis of L. blattarum infection is made on the basis of observation of the characteristic trophozoite in various samples. It is of a great importance to distinguish the protozoon from ciliated respiratory epithelium to avoid wrong positivity. The presented case developed an acute respiratory distress syndrome a short while after taking nivolumab immunotherapy. The morphological features of L. blattarum were demonstrated by examining the bronchoalveolar lavage fluid of the patient under light microscopy. Additionally, URL (https://youtu.be/EQIAsFl6AJY) of a smart-phone based video of trophozoite of this patient was added into this report.

Gastroenteritis caused by single or multiple pathogens remains a major diagnostic challenge for the laboratory, as diagnosis is achieved using different techniques with variable sensitivity and specificity. The aim of this study was to evaluate the EasyScreen™ Enteric Protozoa Detection Kit, a multiplex PCR assay for the detection and identification of the 5 most common protozoan parasites in fecal samples. A total of 632 fecal samples, submitted for routine screening to 2 centers in north-eastern Italy, was included in the study. The results of the molecular assay were compared to those of the standard diagnostic procedures, represented by microscopy and immunoassays. Out of 32 samples testing positive by conventional tools, 31 were detected as concordantly positive using the EasyScreen Kit. Additionally, 91 out of 632 samples only tested positive by the molecular test, therefore increasing the positive detection rate by 275%. Finally, the EasyScreen assay detected 14 co-infections compared to 3 co-infections identified by conventional methods. The EasyScreen Kit provided a rapid and sensitive simultaneous identification of the most important diarrhea-causing protozoa that infect humans. Additionally, this molecular assay presents several advantages compared to conventional tools, such as the standardization and near-total automation of the process. Although critical issues related to the employment of molecular assays are still evident, the system is suitable for clinical parasitological diagnosis as long as it is used in association with conventional tools.

Animals reared in restricted environments are highly susceptible to gastrointestinal infection by helminths and protozoa and therefore zoos are characterized as being parasite-rich environments. Successful implementation of control programs of these parasites in zoo environment depends upon precise and rapid diagnosing of gastrointestinal infections. The aim of this study was to demonstrate the role of the Mini-FLOTAC technique in combination with Fill-FLOTAC for rapidly diagnosing parasitic infections in zoo mammals. Fecal samples were collected from 70 animals in four different zoos located in central and southern Italy. All the samples were analyzed using Mini-FLOTAC in combination with Fill-FLOTAC. Out of the 70 pooled samples examined, 80% (24/30) were positive for at least one parasite. Among the gastrointestinal nematodes, Strongyles were the most frequent (40%), followed by Trichuris spp. (23.3%), Parascaris spp. (13.3%) and Capillaria spp. (3.3%). Among the protozoa, Blastocystis spp., Giardia spp. and Eimeria spp. were detected in 6.6%, 3.3% and 3.3%, respectively. These results show that Mini-FLOTAC in combination with Fill-FLOTAC can be used, not only for rapidly diagnosing parasitic infections in zoo mammals, but also for monitoring control programs in which large numbers of fecal samples need to be examined rapidly and reliably.

Background: Enteric infections are common where public health infrastructure is lacking. This study assesses risk factors for a range of enteric infections among children living in low-income, unplanned communities of urban Maputo, Mozambique., Methods & Findings: We conducted a cross-sectional survey in 17 neighborhoods of Maputo to assess the prevalence of reported diarrheal illness and laboratory-confirmed enteric infections in children. We collected stool from children aged 1-48 months, independent of reported symptoms, for molecular detection of 15 common enteric pathogens by multiplex RT-PCR. We also collected survey and observational data related to water, sanitation, and hygiene (WASH) characteristics; other environmental factors; and social, economic, and demographic covariates. We analyzed stool from 759 children living in 425 household clusters (compounds) representing a range of environmental conditions. We detected ≥1 enteric pathogens in stool from most children (86%, 95% confidence interval (CI): 84-89%) though diarrheal symptoms were only reported for 16% (95% CI: 13-19%) of children with enteric infections and 13% (95% CI: 11-15%) of all children. Prevalence of any enteric infection was positively associated with age and ranged from 71% (95% CI: 64-77%) in children 1-11 months to 96% (95% CI: 93-98%) in children 24-48 months. We found poor sanitary conditions, such as presence of feces or soiled diapers around the compound, to be associated with higher risk of protozoan infections. Certain latrine features, including drop-hole covers and latrine walls, and presence of a water tap on the compound grounds were associated with a lower risk of bacterial and protozoan infections. Any breastfeeding was also associated with reduced risk of infection., Conclusions: We found a high prevalence of enteric infections, primarily among children without diarrhea, and weak associations between bacterial and protozoan infections and environmental risk factors including WASH. Findings suggest that environmental health interventions to limit infections would need to be transformative given the high prevalence of enteric pathogen shedding and poor sanitary conditions observed., Trial Registration: ClinicalTrials.gov NCT02362932., Competing Interests: The authors have declared that no competing interests exist.

Introduction: Intestinal parasitic infections are a major public health problem in inter-tropical areas. The aim of our study was to describe the situation in Mahajanga, Madagascar with a particular focus on two protozoa, Dientamoeba fragilis and Blastocystis sp., Methods: This was a prospective study from February to June 2015. Stool samples from symptomatic hospitalized patients and asymptomatic volunteers were submitted to microscopy and molecular assays in order to detect parasites., Results: A wide panel of intestinal parasites were identified among the 265 included subjects, protozoa being the most prevalent with 72.8% whereas the prevalence of helminths and microsporidia was of 7.9% and 4.5%, respectively. Blastocystis sp. was the most prevalent protozoa (64.5% of the entire cohort) followed by various amoebas (35.5%) and flagellates (27,5%). We only detected subtypes 1, 2 and 3 of Blastocystis sp. Among the patients positive for D. fragilis (9.4%), 23 carried genotype 1 and 1 genotype 2. For the first time, we detected in 4 human stools the DNA of a recently described protozoon, Simplicimonas similis. Interestingly, subjects living in urban areas harbored significantly more different parasitic species than subjects living in rural areas with a correlation between sanitary level of neighborhood and protozoan infection. However, there was no difference in prevalence of digestive symptoms between parasite-free and parasite-infected subjects, except for Giardia intestinalis which had more symptomatic carriers., Discussion: Our study reveals a high overall parasite prevalence, similar to what had been found in 2003 in the same city and to other prevalence studies conducted in Africa. The poor access of the population to sanitary infrastructures may explain this result. Data from our study provide valuable key for sanitation programs and prevention of fecal-related infectious diseases., Competing Interests: The authors have declared that no competing interests exist.

Protozoic infections are prevalent worldwide, particularly in immunosuppressed patients. We reported the case of a patient from the city of Viña del Mar, Chile, a carrier of acute myeloid leukemia in whom an infection by Lophomonas sp. was confirmed by bronchoalveolar lavage. She was treated with antibiotics but died of complications of the underlying disease. There is little literature available on this microorganism. We conclude that Lophomonas sp. should be considered as a diagnostic possibility if protozoa are found in bronchoalveolar lavage of immunosuppressed patients.

Brain-eating amoebae (Acanthamoeba spp., Balamuthia mandrillaris, Naegleria fowleri) have gained increasing attention owing to their capacity to produce severe human and animal infections involving the brain. Early detection is a pre-requisite in successful prognosis. Here, we developed a nanoPCR assay for the rapid detection of brain-eating amoebae using various nanoparticles. Graphene oxide, copper and alumina nanoparticles used in this study were characterized using Raman spectroscopy measurements through excitation with a He-Ne laser, while powder X-ray diffraction patterns were taken on a PANanalytical, X'Pert HighScore diffractometer and the morphology of the materials was confirmed using high-resolution transmission electron microscopy (HRTEM). Using nanoparticle-assisted PCR, the results revealed that graphene oxide, copper oxide and alumina nanoparticles significantly enhanced PCR efficiency in the detection of pathogenic free-living amoebae using genus-specific probes. The optimal concentration of graphene oxide, copper oxide and alumina nanoparticles for Acanthamoeba spp. was determined at 0.4, 0.04 and 0.4 μg per mL respectively. For B. mandrillaris, the optimal concentration was determined at 0.4 μg per mL for graphene oxide, copper oxide and alumina nanoparticles, and for Naegleria, the optimal concentration was 0.04, 4.0 and 0.04 μg per mL respectively. Moreover, combinations of these nanoparticles proved to further enhance PCR efficiency. The addition of metal oxide nanoparticles leads to excellent surface effect, while thermal conductivity property of the nanoparticles enhances PCR productivity. These findings suggest that nanoPCR assay has tremendous potential in the clinical diagnosis of parasitic infections as well as for studying epidemiology and pathology and environmental monitoring of other microbes.

Infections caused by protozoan parasites affect millions of people around the world. Traditionally, diagnosis was made by microscopy, which is insensitive and in some cases not specific. Molecular methods are highly sensitive and specific, but equipment costs and personnel training limit its availability only to specialized centers, usually far from populations with the highest risk of infection. Inexpensive methods that can be applied at the point of care (POC), especially in places with limited health infrastructure, would be a major advantage. Isothermal amplification of nucleic acids does not require thermocyclers and is relatively inexpensive and easy to implement. Among isothermal methods, recombinase polymerase amplification (RPA) is sensitive and potentially applicable at POC. We and others have developed RPA diagnostic tests to detect protozoan parasites of medical importance. Overall, our results have shown high specificity with limits of detection similar to PCR. Currently, the optimization of RPA for use at the POC is under development, and in the near future the tests should become available to detect protozoan infections in the field. In this review we discuss the current status, challenges, and future of RPA in the field of molecular diagnosis of protozoan parasites., (Copyright © 2018 Elsevier B.V. All rights reserved.)

The use of molecular diagnostic techniques in the study of ticks and tick-borne agents has increased dramatically in the last few decades, and while conferring the advantages of rapidity, sensitivity and accuracy, they can also be misused by the research community, who have shown an increasing overreliance on the data they produce, often completely abandoning more traditional approaches such as microscopy and serology. In this short review examples are provided of 'findings' that are apparently at odds with well-established eco-epidemiological knowledge, ranging from descriptions of new species of pathogens to their detection in new hosts and ticks, often immediately incriminated as new reservoir hosts and vector ticks, respectively. Most examples have been taken from the Piroplasmorida or "piroplasm" species as they are often called. Researchers are urged to utilize molecular methods with as much awareness as possible of the system or systems they are investigating, and journal editors and their teams of reviewers should be encouraged to adopt a more critical approach to papers that contain data based solely on molecular methods without due regard to the underlying biology. Explanations for such findings should be offered and discussed., (Copyright © 2018 Elsevier GmbH. All rights reserved.)

Objective: Efficient and easy-to-use DNA extraction and purification methods are critical in implementing PCR-based diagnosis of pathogens. In order to optimize the routine clinical laboratory diagnosis of eukaryotic enteric pathogens, we compare, via quantitative PCR cycle threshold (Ct) values, the efficiency of two DNA extraction kits: the semi-automated EZ1 ® (Qiagen) and the manual QIAamp ® DNA Stool Mini Kit (Qiagen), on six protozoa: Blastocystis spp., Cryptosporidium parvum/hominis, Cyclospora cayetanensis, Dientamoeba fragilis, Giardia intestinalis and Cystoisospora belli and one microsporidia: Enterocytozoon bieneusi., Results: Whereas EZ1 ® (Qiagen) and QIAamp ® DNA Stool Mini Kit (Qiagen) yielded similar performances for the detection of Cryptosporidium spp. and D. fragilis, significant lower Ct values (p < 0.002) pointed out a better performance of EZ1 ® on the five remaining pathogens. DNA extraction using the semi-automated EZ1 ® procedure was faster and as efficient as the manual procedure in the seven eukaryotic enteric pathogens tested. This procedure is suitable for DNA extraction from stools in both clinical laboratory diagnosis and epidemiological study settings.

Clostridium difficile, a Gram-positive spore-forming bacillus, is the most common identifiable etiologic agent of antibiotic-associated diarrhea. The incidence of Clostridium difficile infections among hospitalized children has been increasing across the world. The aim of our study was to evaluate occurrence of Clostridium difficile and some other gastrointestinal pathogens among hospitalized pediatric patients in Georgia, as far as currently statistical data on the topic is very limited in the country. One of the objectives of the study was to test and pilot the real-time Polymerase Chain Reaction diagnostic systems for rapid and simultaneous identification of number of pathogens with a particular emphasis on diarrheal disease diagnostics as these are one of the primary public health priorities in Georgia and worldwide. Cross-Sectional study has been performed on 211 samples collected from 192 pediatric patients. Two pediatric hospitals were involved in the study: M. Iashvili Children's Central Hospital and Tbilisi Children's Clinical Hospital for Infectious Diseases. Laboratory investigations were done in the Clinic NeoLab, Tbilisi, Georgia. Study materials collected for testing were stool samples. Samples were tested by EIA kits (CerTest biotec, Zaragoza, Spain) for presence of A/B toxin according to the manufacturer's instructions. EIA test positive samples were analyzed by home-made multiplex real-time polymerase chain reaction (NeoPCR Diagnostics, NeoLab, Tbilisi, Gerogia) for confirmation of the infection and for simultaneous identification of additional gastrointestinal pathogens including Entamoeba histolitica, Giardia lamblia, Cryptsporidium parvum, Adenovirus, Rotavirus, Norovirus and Astrovirus. All samples were also tested for the presence of the above listed pathogenic agents using the same type EIA kits as for Clostridium difficile described above (CerTestbiotec, Zaragoza, Spain) for presence of the corresponding pathogen. The average age of the study participants was 3.5 years, 56.7% were male and 43.3% were female patients. Presence of Clostridium difficile have been documented in 21 samples out of 211 (10%). Besides the Clostridium difficile, other enteric pathogens have been revealed with the following frequencies: Rotavirus in 12 cases (5.7%), Adenovirus in 11 (5.2%), Giardia lamblia in 10 (4.7%), Astrovirus in 3 (1.4%), Cryptsporidium parvum in 3 (1.4%), Entamoeba histolitica in 2 (0.9%), Norovirus in 2 (0.9%). 49 samples were from out-patient cases (2 samples were positive for Clostridium difficile) and 162 samples were from in-patient cases (19 samples were positive for Clostridium difficile). Clostridium difficile is the frequent pathogenic agent causing diarrheal disease among hospitalized pediatric patients. Development of Clostridium difficile related diarrhea is associated with the antibiotic treatment of pediatric patients hospitalized due to different clinical diagnosis. Targeted early detection of these pathogens is important for the optimal management of diarrheal infection in pediatric patient which will lead to the better clinical outcome and reduction of morbidity rate among hospitalized pediatric patients.

Although microscopic examination of stool samples remains the reference method for the diagnosis of intestinal protozoal infections, these techniques are time-consuming and require operators who are experienced and well trained. Molecular biology seems to offer performances at least equivalent in terms of sensitivity and specificity for certain parasites. This study aimed to compare three multiplex PCR assays on 93 prospectively collected positive stools (prospective cohort) and a panel of 12 more Cryptosporidium-positive samples (Cryptosporidium panel). On the prospective cohort, the sensitivity was 89%, 64% and 41% for Giardia sp. detection for BD Max TM , G-DiaPara TM and RIDA ® GENE, respectively and 75%, 100% and 100% for C. parvum/hominis detection. The sensitivity of the RIDA ® GENE assay for all Cryptosporidium species was 100%, and for D. fragilis 71%. All the techniques obtained the same results for E. histolytica detection, with one positive sample. All species in the Cryptosporidium panel were identified by the RIDA ® GENE PCR. The BD Max TM and G-DiaPara TM assays detected only C. parvum/hominis with the exception of one positive sample for C. meleagridis. No assay showed satisfactory results for all parasites simultaneously, and the DNA extraction seems to be the critical step. More studies are needed to standardize this procedure., (© B. Autier et al., published by EDP Sciences, 2018.)

This protocol is an extension to: Nat. Protoc. 5, 503-515 (2010); doi: 10.1038/nprot.2009.235; published online 25 February 2010The FLOTAC is a sensitive, accurate, and precise technique for the diagnosis of protozoan and helminth infections in humans and animals. However, it requires centrifugation, and hence might be out of reach in resource-constrained settings. As an extension of the original FLOTAC protocol, this protocol describes the Mini-FLOTAC technique, a logical evolution of FLOTAC conceived to perform multivalent, qualitative, and quantitative diagnosis of helminth and protozoan infections in human and animal feces, and urine. This has been found to be of most use in the processing of large numbers of samples with rapid laboratory workup, and for veterinary applications directly on-farm. In addition to the Mini-FLOTAC apparatus, we describe the use of the Fill-FLOTAC, a closed system used to facilitate the performance of the first four consecutive steps of the Mini-FLOTAC technique: fecal sample collection and weighing, homogenization, filtration, and filling of the Mini-FLOTAC chambers. Processing of an individual sample using this protocol requires ∼12 min.

We carried out a cross-sectional study from January to December 2015 on 1,425 inhabitants from a floating population in the Brazilian Amazon (Murinin district, Pará State) to describe the population-based prevalence of tuberculosis (TB) from 2011 to 2014, recent TB contacts (rCts) latently infected with Mycobacterium tuberculosis (LTBI) , the coverage of the local health network, socio-environmental factors, and frequency of intestinal parasitic infection (IPI). We found that the sanitary structure was inadequate, with latrines being shared with other rooms within the same accommodation; well water was the main source of water, and 48% of families had low incomes. The average rate of TB was 105/100, 000 inhabitants per year; one third of TB patients had been household contacts of infected individuals in the past, and 23% of rCts were LTBI. More than half (65%) of 44% of the stools examined (representing 76% of the housing) had IPIs; the highest prevalence was of fecal-oral transmitted protozoa (40%, Giardia intestinalis ), followed by soil-transmitted helminths (23%). TB transmission may be related to insufficient disease control of rCts, frequent relocation, and underreporting. Education, adopting hygienic habits, improving sanitation, provision of a treated water supply and efficient sewage system, further comprehensive epidemiological surveillance of those who enter and leave the community and resources for basic treatment of IPIs are crucial in combating the transmission of these neglected diseases.

Background: Bronchopulmonary disease caused by flagellated protozoa infection (BPFP) is thought to be rare in children but may be an emerging or underestimated disease, especially in developing countries., Methods: In this study, we retrospectively reviewed records of 15 patients who were presented with a cough, wheezing or bronchopulmonary disease of unknown causes during admission, and patients who were finally diagnosed with BPFP from January 2014 to January 2015 were enrolled. Protozoa were observed in bronchoalveolar lavage fluid by light microscopy., Results: A total of 15 pediatric cases (11 male and 4 female individuals, from 1 year 8 months to 12 years 1 month of age) with flagellated protozoa infection diagnosed by light microscopy were recruited. The course of the disease at the time of diagnosis was from 10 days to 11 months. Patients presented with a fever (N = 9), cough (N = 11), wheezing (N = 5) and chest pain (N = 5). Laboratory data showed elevated peripheral blood leucocytes (N = 6), eosinophilic granulocytes (N = 3), C-reactive protein (N = 5) and immunoglobulin E (N = 3). Bronchoscopy revealed a mucus plug (N = 3) and bronchiectasis (N = 1). Lung computed tomography results indicated ground-glass opacification (N = 2), atelectasis (N = 3), bronchiectasis (N = 1), bronchial wall thickening (N = 3) or nodular opacity (N = 6, including 1 case of pulmonary embolism). All children responded to metronidazole for a 2- to 5-week treatment period., Conclusions: Patients with BPFP often have a chronic or recurrent course and present with recurrent fever, cough, wheezing and chest pain. Chest imaging may reveal ground-glass opacification, atelectasis, bronchiectasis or nodular opacity (including pulmonary embolism). BPFP responds favorably to metronidazole treatment.

The objective of this study was to standardize the high-resolution melting method for identification and discrimination of Toxoplasma gondii, Sarcocystis spp., Neospora spp., and Cryptosporidium spp. by amplification of 18S ribosomal DNA (rDNA) using a single primer pair. The analyses were performed on individual reactions (containing DNA from a single species of a protozoan), on duplex reactions (containing DNA from two species of protozoa in each reaction), and on a multiplex reaction (containing DNA of four parasites in a single reaction). The proposed method allowed us to identify and discriminate the four species by analyzing the derivative, normalized, and difference melting curves, with high reproducibility among and within the experiments, as demonstrated by low coefficients of variation (less than 2.2% and 2.0%, respectively). This is the first study where this method is used for discrimination of these four species of protozoa in a single reaction.

Histomonas meleagridis is a flagellate protozoan organism that can cause severe necrotizing typhlitis and hepatitis in gallinaceous birds. Peafowl ( Pavo spp.) have been shown to be susceptible to histomoniasis in experimental settings, but there are few reports of natural histomoniasis in this species. A retrospective study of the archived cases at 2 veterinary diagnostic laboratories in the United States yielded 5 cases of peafowl with gross and histologic findings characteristic of histomoniasis. Lesions included bilateral, transmural fibrinonecrotic typhlitis and multifocal necrotizing hepatitis with associated trophozoites morphologically consistent with H. meleagridis. There was no evidence of Heterakis gallinarum infestation in the studied cases. DNA was extracted from formalin-fixed, paraffin-embedded liver and ceca from all 5 cases and was analyzed using multiple sets of primers with subsequent sequencing and genotyping. Four samples were positive for H. meleagridis, and 1 sample was positive for both H. meleagridis and Tetratrichomonas gallinarum. These results confirm that peafowl develop clinical disease similar to that described previously in other gallinaceous birds infected by H. meleagridis. The role of T. gallinarum remains unknown and further research is necessary to elucidate its role, if any, in the pathogenesis of the observed lesions.

Detection of intestinal parasites from fecal samples is routinely performed by direct wet mount examination. This method requires skilled personnel, and it is time consuming. The aim of this work is to demonstrate the usefulness of the newer automated urinary sediment analyser sediMAX 2 for a fast detection of intestinal protozoa in stool samples. A total of 700 consecutively preserved samples consisting of 70 positives and 630 negatives were analyzed. SediMAX 2 takes digital images of each sediment sample, and analysis was conducted using a dilution of stool specimens, allowing determination of typical morphology. Compared to manual microscopy, sediMAX 2 showed sensitivity and specificity of 100 % in the detection of intestinal parasites, as also recently demonstrated for sediMAX 1. However, all clinically important human protozoa were detected using only 15 images for each specimen, compared to 30 images required in sediMAX 1 analysis. Moreover, changing manually the focus, it is possible to carry out a discrimination between morphologically identical Entamoeba complex members, including the pathogenic E. histolytica and the non-pathogenic E. dispar, E. moshkovskii and E. Bangladeshi, from the non-pathogenic Entamoeba coli based on the number of nuclei present in the cells. This study presents sediMAX 2 as an automatic aid to traditional microscopy.

Introduction: The diagnosis of intestinal parasitic infections depends on the parasite load, the specific gravity density of the parasite eggs, oocysts or cysts, and the density and viscosity of flotation or sedimentation medium where faeces are processed., Objective: To evaluate the concordance between zinc sulphate flotation and centrifugal sedimentation in the recovery of parasites in faecal samples of children., Materials and Methods: Faecal samples of 330 children from day care centers were evaluated by zinc sulphate flotation and centrifugal sedimentation techniques. The frequencies of detection of parasites by each method were determined and the agreement between the diagnostic techniques was evaluated using the kappa index, with 95% confidence intervals., Results: The faecal flotation in zinc sulphate diagnosed significantly more cases of Trichuris trichiura infection when compared to centrifugal sedimentation (39/330; 11.8% vs. 13/330; 3.9%, p<0.001), with low diagnostic concordance between methods (kappa=0.264; 95% CI: 0.102-0.427). Moreover, all positive samples for Enterobius vermicularis eggs (n=5) and Strongyloides stercoralis larvae (n=3) were diagnosed only by zinc sulphate. No statistical differences were observed between methods for protozoa identification., Conclusions: The results showed that centrifugal flotation in zinc sulphate solution was significantly more likely to detect light helminths eggs such as those of T. trichiura and E. vermicularis in faeces than the centrifugal sedimentation process.

Infection with pathogenic intestinal protozoa. as Giardia lamblia,, Entamoeba histolytica and Cryptosporidium parvum cause considerable gastrointestinal morbidity, malnutrition and mortality worldwide, especially among young children in developing countries. The present study was carried out on 71 cases (44 males & 27 females) chosen from Pediatric and Internal Medicine Inpatient and Outpatient Clinics of Zagazig University Hospitals, complaining of different gastrointestinal troubles with an age range of 6-60 years. Also, 20 apparently healthy individuals (11 males & 9 females) cross matched were considered as a control negative group. All stool samples were examined by direct wet smears, concentration techniques, staining of the smears using trichrome stain and Modified Ziehl-Neelsen method. Copro-antigen detection in faecal sample was by using quick immunochromatographic test. A total of 71 cases suffering from different GIT manifestations showed G. lamblia (30.7%), Cryptosporidium parvum (19.8%), EntamoebahistolyticalE. dispar (11%) and mixed infection of three protozoa (6.6%). However, by copro-antigen G. lamblia was positive in (31.8%) of C. parvum in (20.9%); E. histolytica/E. dispar in (11%) of cases. Immunochromatography/copro-antigen test recorded sensitivity and spec- ificity of (100%) and (96.6%) respectively in G. lamblia detection. For C. parvum, sensitivity was (100%) and specificity was (97.1%) while for E. histolytica/E. dispar. sensitivity and specificity were (100%) for both. Immunochromatographic assay proved to be simple, easy and useful in confirming absence or the presence of intestinal protozoan infection in clinically suspected cases with negative stool examination.

Background: Parasitic infections are still a significant health problem in rural areas in developing countries including Iran. There is no recent population-based data about the prevalence of human intestinal parasites in most rural areas of Iran. The current study aimed to determine the prevalence of intestinal protozoan infection in inhabitants of rural areas of Boyer-Ahmad district, Southwestern Iran., Methods: A total of 1025 stool samples were collected from the inhabitant of 50 randomly selected villages in Boyer-Ahmad Township. The stool samples were evaluated by parasitological methods including, direct wet-mounting, formalin ethyl acetate concentration, zinc sulfate floatation, and Trichrome permanent stain for detection of protozoan infections. Diarrheic samples were further evaluated with a modified Ziehl-Neelsen staining method for detection of coccidian parasites., Results: The prevalence of both pathogenic and nonpathogenic intestinal parasites in the population was 37.5% (385 out of 1025 cases), some individual with multiple infections. Giardia lamblia was detected in 179 (17.46%), Blastocystis hominis in 182 (17.76%), Entamoeba histolytica/dispar in 9 (0.87%), Endolimax nana in 216 (21.07%), Entamoeba coli in 151 (14.73%), Ioedamoeba butschlii in 45 (4.39%), Chillomastix mesnili in 22 (2.14%), Trichomonas hominis in 2 (0.19%) and Dientamoeba fragillis in 2 (0.19%) of cases. Multivariate logistic regression revealed significant associations between protozoan infection (pathogenic protozoa) and contact with animals (OR yes/no = 2.22, p < 0.001) and educational status (OR higher/illiterate = 0.40, P = 0.01)., Conclusion: Findings of this study demonstrated that protozoan infection rate in rural areas of southwestern Iran is still high and remained as a challenging health problem in these areas.

In resource-limited settings, the scarcity of skilled personnel and adequate laboratory facilities makes the differential diagnosis of fevers complex [1-5]. Febrile illnesses are diagnosed clinically in most rural centers, and both Rapid Diagnostic Tests (RDTs) and clinical algorithms can be valuable aids to health workers and facilitate therapeutic decisions [6,7]. The persistent fever syndrome targeted by NIDIAG is defined as presence of fever for at least one week. The NIDIAG clinical research consortium focused on potentially severe and treatable infections and therefore targeted the following conditions as differential diagnosis of persistent fever: visceral leishmaniasis (VL), human African trypanosomiasis (HAT), enteric (typhoid and paratyphoid) fever, brucellosis, melioidosis, leptospirosis, malaria, tuberculosis, amoebic liver abscess, relapsing fever, HIV/AIDS, rickettsiosis, and other infectious diseases (e.g., pneumonia). From January 2013 to October 2014, a prospective clinical phase III diagnostic accuracy study was conducted in one site in Cambodia, two sites in Nepal, two sites in Democratic Republic of the Congo (DRC), and one site in Sudan (clinicaltrials.gov no. NCT01766830). The study objectives were to (1) determine the prevalence of the target diseases in patients presenting with persistent fever, (2) assess the predictive value of clinical and first-line laboratory features, and (3) assess the diagnostic accuracy of several RDTs for the diagnosis of the different target conditions., Competing Interests: The authors have declared that no competing interests exist.

The implementation of polymerase chain reaction (PCR)-based diagnostics of intestinal protozoa has led to higher sensitivity and (subtype) specificity, more convenient sampling, and the possibility for high-throughput screening. PCR for routine detection of human intestinal protozoa in fecal samples is used by an increasing number of clinical laboratories. This paper discusses the recent developments in the diagnosis of intestinal protozoa, with an emphasis on PCR-based diagnostics. Although many reviews have described the technical aspects of PCR-based diagnostics, this review focuses on the clinical consequences that result from the shift from microscopic toward PCR-based diagnostics. Despite its undisputed superiority, the use of PCR comes with challenges that clinicians should be aware of., (© 2016 Journal of Gastroenterology and Hepatology Foundation and John Wiley & Sons Australia, Ltd.)

Waterborne diseases associated with polluted recreational and potable waters have been documented for more than a century. Key microbial protozoan parasites, such as Cryptosporidium and Giardia, are causative agents for gastrointestinal disease worldwide. Although not a first-line diagnostic approach for these diseases, medical imaging, such as radiography, computed tomography, magnetic resonance imaging, ultrasonography, and nuclear medicine technologies, can be used to evaluate patients with long-term effects. This article describes protozoan pathogens that affect human health, treatment of common waterborne pathogen-related diseases, and associated medical imaging., (©2016 American Society of Radiologic Technologists.)

Intestinal parasites are a major public health problem in developing countries, most prevalent in areas where sanitation is poor and the population's hygiene is inadequate. They affect people of all ages, although school-age children are the most susceptible. In this study, we investigated the presence of intestinal parasites in the shared environments of a public school in the town of Diamantina, Minas Gerais State. From December 2012 to February 2013, samples were collected for three months (once a month) by using the Graham method (1941), in duplicate, by affixing a 6 x 5 cm clear tape, six times in each collection site, in a space of about 30 cm2. Then, each tape was positioned longitudinally on a microscope slide and the identification of the biological forms of the parasites was performed with the aid of a 40X objective from an optical microscope. Eleven sites were selected for sampling. Cysts of Entamoeba coli were the most frequently found in this study (50%), followed by Hymenolepis diminuta eggs (27.6%), Iodamoeba butschllii cysts (5.6%), Ascaris lumbricoides eggs (5.6%), Taenia species eggs (5.6%) and hookworm eggs (5.6%). The highest positivity rates were found in the samples drawn from the cafeteria's eating table. The results have indicated the need to improve the cleaning in the school environments, as well as the development of educational practices that may help in the preservation of public health.