The Distribution of Intestinal Amoebae in Wild Long-Tailed Macaques (Macaca fascicularis) in Sabang City, Aceh Province, Indonesia

Amoeba that infects primates can also be found in humans and can be zoonotic. This study aims to identify the distribution of intestinal amoeba in the wild Long-Tailed Macaque (LTM) group located at Cot Murong, Kebun Mangga, and Tugu Zero Kilometer Monument (IZKM) in Sabang City, Aceh Province, Indonesia. A total of 177 stool samples from 59 tails from the three LTM groups were examined microscopically. The results of the examination confirmed seven species of intestinal amoeba, three of which belonged to the genus Entamoeba, namely the Entamoeba histolytica/dispar/moshkovskii complex, Entamoeba coli, Entamoeba pölecki, and four other species from different genera, namely Iodamoeba butschlii, Blastocystis sp., Endolimax nana and Dientamoeba fragilis. The prevalence of intestinal amoeba in wild LTM in Sabang City was 57.6% (102/177). Dientamoeba fragilis was only found in the LTM group of Kebun Mangga. Entamoeba coli dominated the highest prevalence in all LTM groups. The LTM group located around Cot Murong and IZKM had a higher concentration of infection with intestinal amoeba than the Kebun Mangga location. The statistical tests showed a significant difference between the proportions of Iodamoeba butschlii, Endolimax nana and Dientamoeba fragilis, both found in the LTM Cot Murong, Kebun Mangga, or IZKM groups. This study is the first to report amoebic infection prevalence in the wild LTM group in Sabang City, Aceh Province. The pathogenic amoeba that cannot be distinguished microscopically from morphologically identical amoebae needs to be identified molecularly to identify variants and their potential as zoonotic agents. HIGHLIGHTS Intestinal amoeba species found in fecal samples of the wild long-tailed macaques group of Sabang City, Aceh Province, Indonesia, can also be found in human stool samples As many as three of the seven identified amoeba species, namely Entamoeba histolytica, Dientamoeba fragilis, and Blastocystis sp. is a pathogen that can cause diarrhea and colitis in humans and non-human primates The prevalence of amoeba infection in wild macaques in Sabang City is in the high category and tends to increase the incidence of infection in the macaque group, which has more susceptible individuals, namely infants, children, and females GRAPHICAL ABSTRACT

Investigation of neglected protists Blastocystis sp. and Dientamoeba fragilis in immunocompetent and immunodeficient diarrheal patients using both conventional and molecular methods

Introduction The clinical significance of Blastocystis sp. and Dientamoeba fragilis in patients with gastrointestinal symptoms is a controversial issue. Since the pathogenicity of these protists has not been fully elucidated, testing for these organisms is not routinely pursued by most laboratories and clinicians. Thus, the prevalence of these organisms and the subtypes of Blastocystis sp. in human patients in Turkey are not well characterized. This study aimed to determine the prevalence of Blastocystis sp. and D. fragilis in the diarrheic stool samples of immunodeficient and immunocompetent patients using conventional and molecular methods and to identify Blastocystis sp. subtypes using next generation sequencing. Material and methods Individual stool specimens were collected from 245 immunodeficient and 193 immunocompetent diarrheic patients between March 2017 and December 2019 at the Gazi University Training and Research Hospital in Ankara, Turkey. Samples were screened for Blastocystis sp. and D. fragilis by conventional and molecular methods. Molecular detection of both protists was achieved by separate qPCRs targeting a partial fragment of the SSU rRNA gene. Next generation sequencing was used to identify Blastocystis sp. subtypes. Results The prevalence of Blastocystis sp. and D. fragilis was 16.7% and 11.9%, respectively as measured by qPCR. The prevalence of Blastocystis sp. and D. fragilis was lower in immunodeficient patients (12.7% and 10.6%, respectively) compared to immunocompetent patients (21.8% and 13.5%, respectively). Five Blastocystis sp. subtypes were identified and the following subtype distribution was observed: ST3 54.4% (n = 37), ST2 16.2% (n = 11), ST1 4.4% (n = 3), ST6 2.9% (n = 2), ST4 1.5% (n = 1), ST2/ST3 11.8% (n = 8) and ST1/ST3 8.8% (n = 6). There was no statistically significant difference in the distribution of Blastocystis sp. subtypes between immunocompetent and immunodeficient patients. Conclusion and recommendation Our findings demonstrated that Blastocystis sp. and D. fragilis are commonly present in immunocompetent and immunodeficient patients with diarrhea. This study is the first to use next generation sequencing to address the presence of Blastocystis sp. mixed subtypes and intra-subtype variability in clinical samples in Turkey.

Fecal screening of Dientamoeba fragilis: relative efficacy of permanent smear and culture

Dientamoeba fragilis (D. fragilis) is an enteric trichmonad protozoan parasite that remains obscure and neglected. The aim of this study is to detect D. fragilis as a neglected pathogen in children aged 6-12 years old complaining of gastrointestinal illness by stool culture and light microscopy with comparison between the results of both techniques. A total of 100 fresh stool samples were included in this current study. All specimens were subjected to microscopic examination using iron- hematoxylin stained stool smears and stool culture using a Loeffler’s culture medium. Culture detected 2 positive stool samples (2%) while microscopy detected (1%). Sensitivities of culture and microscopy were 100% and 50% respectively. Specificity of culture and microscopy were 100% and 95% respectively. There is a moderate agreement between culture and microscopy (K = 0.4). In conclusion, culture had a high performance compared to microscopy in the diagnosis of D. fragilis infection. Culture can be applied for routine diagnosis for the detection of D. fragilis in clinical samples.

A Novel Culprit in a Patient with Budd-Chiari Syndrome

Budd-Chiari syndrome (BCS) is an uncommon illness that is characterized by obstruction of hepatic venous outflow. Patients typically present with nausea, vomiting, and abdominal pain, which can further progress into signs associated with liver failure, including jaundice, encephalopathy, and coagulopathy. The most common causes of BCS include pathologies that induce portal vein thrombosis, such as myeloproliferative disorders, malignancy, and acquired hypercoagulable states. In this case report, a patient who presented with abdominal pain and distention is diagnosed with BCS caused by an unusual etiology. He was found to have significant eosinophilia, prompting additional evaluation for parasitic infections. Using stool diagnostics/studies, he was found to have <i>Dientamoeba fragilis</i> trophozoites. The patient was treated with enoxaparin, warfarin, and metronidazole with a resolution of his symptoms. This case outlines a novel cause of BCS as well as the proposed mechanism of <i>Dientamoeba fragilis</i> induction of BCS.